®l|?  E  B.  Bill  iCtbrara 

Nortlj  (Earaltna  i'talp 


^a 


This  book  was  presented  by 

MILTON  M.  LEONARD,  D.V.M. 

TO  THE 

VETERINARY  MEDICAL  LIBRARY 


This  book  is  due  on  the  date  indicated 
below  and  is  subject  to  an  overdue 
fine  as  posted  at  the  circulation  desk. 


EXCEPTION:  Date  due  will  be 
earlier  if  this  item  is  RECALLED. 


THE 


Examination    of    the  Urine 


OF  THE 


HORSE  AND  MAN 


PIERRE  A.  FISH,  D.Sc,  D.V.M. 

PROFESSOR    OF    VETERINARY    PHYSIOLOGY 

NEW  YORK  STATE  VETERINARY  COLLEGE 

CORNELL  UNIVERSITY 


SECOND  EDITION 


PUBLISHED    BY 

CARPENTER   &   CO. 

ITHACA,    N.  Y. 

191 1 


COPYRIGHT,    1906 


PIERRE    A.    FISH 


PREFACE. 


This  manual  represents  the  published  form  of  mimeographed 
sheets,  which  the  writer  has  used,  for  some  years  past,  in  giving  in- 
struction to  his  veterinary  students.  Data  relating  to  the  urine  of  the 
horse  is  not  abundant  and,  unfortunately,  some  of  the  tests  which  may 
be  standard  for  human  urine  are  not  reliable  for  that  of  the  horse  and, 
therefore,  require  modification.  In  order  to  emphasize  this  fact,  the 
writer  has  employed  the  comparative  method  and  asks  the  students  to 
examine  their  own  urine  along  with  that  of  the  horse  and  note  care- 
fully the  differences  in  the  results. 

The  importance  of  urine  examination  for  diagnosis  and  prognosis 
in  human  medicine  is  too  well  known  to  require  emphasis  here;  but, 
notwithstanding  certain  difficulties  in  the  way  of  its  application  in 
veterinary  medicine,  the  writer  believes  that  there  is  no  important  rea- 
son why  just  as  much  valuable  information  may  not  be  derived,  in  cer- 
tain cases,  from  the  urine  of  the  horse  by  an  up-to-date  veterinarian,  as 
the  physician  obtains  from  the  urine  of  his  patients. 

Simplification  of  methods,  without  too  great  a  sacrifice  in  accuracy, 
is  essential  for  satisfactory  examination, — especially  by  veterinarians. 
The  writer  realizes  that  only  a  short  step  has  been  taken  in  this  direc- 
tion, but  hopes  that  time  will  lighten  the  difficulties. 

P.  A.  F. 

October,  1906. 

PREFACE  TO  THE  SECOND  EDITION. 

In  the  present  edition  some  changes  have  been  made  in  the  text 
and  new  material  added  with  the  hope  of  bringing  the  manual  more 
completely  up  to  date.  There  is  still  much  to  be  done  in  investigating 
and  simplifying  the  tests  applicable  to  the  urine  of  the  domesticated 
animals  and  it  is  to  be  hoped  that  more  research  may  be  developed 
along  this  line. 

P.  A.  F. 
November,  1911. 


TABLE  OF  CONTENTS. 


CHAPTER  I. 

The  Secretion  of  Urine pp.     7-12 

CHAPTER  n. 

Quantity,  Color,  Transparency,  Consistency,  Reaction, 

Degree  of  Acidity,  Specific  Gravity pp.  12-19 

CHAPTER  m. 

Qualitative     tests:     Inorganic     Constituents,     Water, 

Chlorides,  Sulphates,  Phosphates,  Carbonates pp.  19-24 

CHAPTER  IV. 

Organic  Constituents:  Urea,  Uric  Acid,  Hippuric 
Acid,  Creatinin,  Mucus,  Indican,  Oxalic  Acid, 
Acetone,  Urobilin,  Leucin,  Tyrosin,  Phenol pp.  24-33 

CHAPTER  V. 

Abnormal  Substances  in  the  Urine:  Albumin,  Sugar,  .pp.  33-41 

CHAPTER  VI. 

Bile,  Blood,  Melanine pp.  41-45 

CHAPTER  VII. 

Quantitative  Analysis,  Centrifugal  Method:  Phos- 
phates, Chlorides,  Sulphates,  Uric  Acid,  Uricom- 
eter.  Urea,  Albumin,  Saccharometer,  Sugar,  Ehr- 
lich's  Diazo-reaction pp.  45-56 

CHAPTER  VIII. 

Volumetric  Methods :  Chlorides,  Phosphoric  Acid,  Sul- 
phuric Acid,  Relation  of  Constituents  in  Normal 
Urine   PP-  56-60 

CHAPTER  IX. 

Chemical  Examination  of  Urinary  Deposits,  Acid, 
Urate  of  Soda,  Uric  Acid,  Cystine,  Pus,  Blood,  Al- 
kaline Urates,  Triple  Phosphates,  Calcium  Oxalate, 
Calcium  Carbonate,  Pus,  Blood pp.  60-63 

CHAPTER  X. 

Microscopical  Examination  of  Urine,  Unorganized  Sedi- 
ments: Uric  Acid,  Acid  Urate  Soda,  Oxalate  of 
Lime,  Hippuric  Acid,  Calcium  Sulphate,  Calcium 
Phosphate,  Triple  Phosphate,  Urate  of  Ammonium, 
Cystin,  Leucin,  Tyrosin.  Organized  Sediments: 
Epithelial  cells;  Glycogen  cells,  Amjdoid  bodies, 
Hyaline,  Granular,  Epithelial,  Fat,  Hemorrhagic 
and  Waxy  casts ;  Cylindroids ;  Blood  and  Pus  cor- 
puscles ;  Spermatozoa ;  Mucus ;  Bacteria pp.  63-73 

Form  for  Urine  Examinations,  Procedure  in  Examin- 
ing a  sample  of  urine PP.  74-77 

Appendix,  Formulae  for  Reagents pp.  77-80 


URINE  ANALYSIS. 


APPARATUS   FOR  THE   LOCKER. 


1  dozen  test  tubes,  6  inch 

1  dozen  test  tubes,  5  inch 

1  Minim  pipette 

1  Beaker,  10  oz. 

1  Graduate,  30  cc. 

1  Graduate,  250  cc. 

1  Flasli 

1  Funnel,  lYz  inch 

1  Funnel,  3  inch 

1  Watch  glass 

1  Evaporating  dish,  S  oz. 

1  Urinometer 

1  Glass  rod 

1  Thermometer 

1  Crucible,  8  cc. 


1  Piece  wire  gauze 

1  Piece  absorbent  cotton 

1  Box  matches 

1  Test  tube  brush 

1  Test  tube  rack 

1  Test  tube  holder,  wire 

1  Tripod 

1  Piece  muslin 

1  Pack  filter  papers,  3  inch 

1  Pack  filter  papers,  6  inch 

1  Sponge 

1  Clay  triangle 

2  Tin  cans 

1  Copper  water  bath 
1  Towel 


Special  apparatus,  not  found  in  the  locker,  may  be  obtained,  when 
needed,  by  handing  an  order  for  it  to  one  of  the  assistants. 


Kidney. 


1.  Cut  portion  of  the  Kidney.  2.  Pyramid.  3.  Papilla  of  the  Pyramid. 
4.  Glomerulus,  enclosed  in  Bowman's  Capsule.  5.  Bowman's  Capsule.  6. 
Convoluted  portion  of  Renal  Tubule.  7.  Loop  of  Henle.  8.  Collecting 
Tubule.  9.  Capillary  net  work, — second  set;  the  first  set  is  the  Glom- 
erulus. 10.  Openings  of  Collecting  Tubules  on  Papillae,  from  which  the 
Urine  drops  into  the  Pelvis  of  the  Kidney. 


THE  SECRETION  OF  URINE. 


I. 

The  blood  supply  to  the  kidneys  is  an  important  consider- 
ation in  the  secretion  of  urine.  At  the  outset,  one  is  impressed 
by  the  fact  that  a  relatively  large  artery  supplies  a  relatively 
small  gland.  The  blood  pressure  of  the  renal  artery  is  nearly, 
if  not  quite,  as  great  as  that  of  the  abdominal  aorta,  which 
means  that  about  as  much  pressure  is  required  to  force  the 
blood  through  the  kidneys  as  is  required  to  send  the  blood 
through  the  pedal  extremities.  The  renal  vein  is  also  a  rela- 
tively large  vessel  as  compared  with  the  size  of  the  kidney  or 
with  the  efferent  vessels  of  other  glands.  The  pressure  within 
the  renal  vein  is  very  low,  practically  as  low  as  that  in  the 
posterior  vena  cava  with  which  it  connects. 

The  artery,  after  entering  the  kidney,  breaks  up  into 
branches  which  pass  between  the  pyramids.  At  the  junction 
of  the  cortex  and  medulla  these  branches  form  arches  in  the 
substance  of  the  kidneys  and  from  these  arches  branches  run 
outward  into  the  cortex  to  supply  the  glomeruli ;  other  branches 
pass  inward  to  supply  the  pyramids.  Each  glomerulus  has 
its  afferent  and  efferent  vessel  and  of  these  the  efferent  is 
smaller. 

On  issuing  from  the  capillaries  of  the  glomerulus,  the 
efferent  vessel  soon  breaks  up  into  a  second  set  of  capillaries 
which  supplies  the  uriniferous  tubules ;  so  that  in  this  arrange- 
ment the  blood  goes  first  to  the  glomeruli  and  later  supplies 
the  tubules.  The  blood  from  the  second  set  of  capillaries  is 
finally  gathered  up  by  small  vessels  which  unite  to  form  ulti- 
mately the  renal  vein.  The  glomeruli  and  the  uriniferous 
tubules  are  the  portions  of  the  kidney  actively  concerned  in 
the  production  of  its  secretion — the  urine. 

The  following  points  are  therefore  worthy  of  special 
notice:  in  the  malpighian  body  the  arterial  blood  in  the  glom- 
erular capillaries  is  separated  from  the  inside  of  the  capsule 
of  Bowman  by  a  thin  layer  of  flattened  epithelial  cells  only; 
two  sets  of  capillaries  exist,  one  set  forming  the  glomeruli  and 
the  other  supplying  the  uriniferous  tubules,  the  blood  supply- 


8 


ing  the  tubules  must  have  first  passed  through  the  glomeruli, 
and  is  therefore  more  concentrated;  in  certain  portions  of  the 
tubules  short  cylindrical  epithelial  cells  are  found  which  are 
comparable  to  true  secreting  cells  found  in  other  glands;  and 
finally,  the  smaller  size  of  the  efferent  as  compared  with  the 
afferent  vessel  fulfills  a  condition  which  retards  the  flow  of 
blood  through  the  glomerulus. 

The  conditions  are  favorable  for  a  high  and  variable 
pressure  in  the  glomerulus  and  for  a  lower  and  more  constant 
pressure  in  the  second  set  of  capillaries  around  the  uriniferous 
tubules.  On  account  of  the  resistance  offered  by  a  double 
system  of  capillaries  the  blood  pressure  in  the  kidneys  is  kept 
relatively  high. 

The  changes  in  blood  pressure  may  be  observed  upon  the 


Fig.  1. 

1.  Artery.  2.  Glomerulus.  3.  Capsule  of  Bowman.  4.  Convoluted 
portion  of  Tubule.  5.  Capillary  net  work.  7.  Loop  of  Henle.  8.  Collect- 
ing Tubule.    9.  Opening  of  Tubule  on  Papilla. 


kidney  itself  by  means  of  an  apparatus  known  as  the  onco- 
meter. With  each  rise  in  the  blood  pressure  the  kidney  swells, 
with  each  fall  in  pressure  it  contracts,  and  a  tracing  can  be 
obtained  very  similar  to  an  ordinary  blood  pressure  tracing. 
The  glomerulus  suspended  in  its  capsule  is  also  influenced  by 
these  changes.  AVhen  the  capillaries  are  engorged  with  blood 
the  glomerulus  fills  the  capsule,  when  collapsed  there  is  an 
'  evident  space  between  the  membranes  of  the  glomerulus  and 
the  capsule. 

In  1842  Bowman  advanced  a  theory  relating  to  the  pro- 
duction of  urine  which,  while  recognizing  to  some  extent  cer- 
tain physical  factors,  also  brought  out  the  view  that  the  urine 
is  a  secretion. 

In  1844  Ludwig  advanced  the  theory,  sometimes  referred 
to  as  the  mechanical  theory,  in  which  only  physical  processes 
were  involved,  i.  e.,  filtration,  diffusion  and  osmosis.  It  is  true 
that  in  the  capillaries  of  the  glomerulus  there  is  a  high  resist- 
ance because  of  the  smaller  size  of  the  efferent  vessel,  there  is, 
therefore,  a  higher  degree  of  pressure  in  the  glomerulus  than 
in  the  capsule  in  which  it  is  suspended.  This  inequality  of 
pressure  is  favorable  to  filtration.  As  the  result  of  any  filtra- 
tion from  the  glomerulus  some  of  the  water  of  the  blood  with 
a  certain  proportion  of  dissolved  substances  would  pass  into  the 
beginning  of  the  uriniferous  tubule.  The  effect  of  such  a 
filtration  would  render  the  blood  remaining  in  the  glomerulus 
and  second  set  of  capillaries  more  concentrated,  and  in  the 
second  set  of  capillaries  in  connection  with  the  uriniferous 
tubules  the  essential  elements  of  an  osmometer  would  be 
obtained, — an  animal  membrane  formed  by  the  delicate  wall 
of  the  capillary  and  the  wall  of  the  tubule,  upon  one  side  of 
which  there  is  a  dense  fluid,  the  blood,  and  upon  the  other  a 
vt^eak  saline  solution,  conditions  which  are  favorable  to  the 
processes  of  osmosis  and  diffusion. 

In  this  theory,  as  a  result  of  the  interchange  of  these 
fluids,  some  of  the  water  passes  from  the  tubule  to  the  blood, 
Tiiaking  it  less  concentrated,  and  the  products  of  retrogressive 
metamorphosis, — urea  and  other  organic  substances, — pass 
from  the  blood  to  the  tubule,  forming  urine.  An  objection  in 
connection  with  the  diffusion  of  urea  has  been  raised  in  that 


10 


it  is  a  well-known  fact  that  the  urine  contains  a  much  greater 
amount  of  urea  than  does  the  blood  and  that  it  would  be  con- 
trary to  the  laws  of  osmosis  and  diffusion  for  a  fluid  weak  in 
urea  (the  blood)  to  pass  through  a  membrane  a  substance 
which   has   accumulated   in   greater   amount   in   a   fluid    (the 

urine)  on  the  other  side  of 
that  membrane.  It  should  also 
be  remembered  that  the  blood 
is  constantly  moving  and  pre- 
sumably the  fluid  in  the  tu- 
bules is  doing  the  same.  In 
other  words,  it  would  seem 
that  if  the  conditions  indicate 
an  osmometer,  it  varies  from 
experimental  ones  in  that  the 
fluids  on  either  side  of  the 
membrane,  are  moving  in  a 
definite  direction.  The  pro- 
teid  constituents  do  not  nor- 
mally leave  the  blood  on  ac- 
count of  their  well-known  in- 
disposition to  osmosis. 
One  important  fact,  however,  remains  unaccounted  for  in 
Ludwig's  mechanical  theory,  and  that  is,  if  the  uriniferous 
tubules  are  stripped  of  their  epithelial  cells,  as  they  often  are 
in  disease,  urea  and  some  other  nitrogenous  products  are  no 
longer,  or  only  imperfectly  eliminated,  and  become  stored  up 
in  the  blood  and  produce  the  condition  known  as  uremia, 
although  the  conditions  of  an  osmometer  remain.  It  must, 
therefore,  be  admitted  that  there  is  some  direct  or  elaborating 
action  on  the  part  of  the  epithelium  as  originally  suggested 
by  Bowman,  although  under  normal  conditions,  transudation, 
diffusion  and  osmotic  processes  may  occur  coincidently. 

The  theoretical  conclusions  of  Bowman  have  been  con- 
firmed and  extended  by  the  practical  researches  of  Heidenhain, 
who  injected  indigo-carmine  into  the  blood  of  animals  and 
found  that  it  was  promptly  removed  by  the  kidneys.  These 
organs  were  removed  at  suitable  intervals  after  the  injection 


Fig.  2. 
1.  Artery.  2.  Afferent  Vessel.  3. 
Glomerulus.  4.  Capsule  of  Bow- 
man. 5.  Efferent  Vessel.  6.  Capil- 
lary network.  7.  Uriniferous  Tub- 
ule.   8.  Vein. 


11 

and  carefully  examined  under  the  microscope.  In  no  instance 
did  he  find  any  of  the  indigo  in  the  capsules  of  Bowman,  but 
it  was  found  abundantly  in  the  cells  lining  certain  portions  of 
the  tubules,  and  in  the  lumen  of  the  tubules  near  these  cells. 
Similar  experiments  with  the  urate  of  sodium  showed  that  it 
likewise  was  secreted  at  the  same  place  and  in  the  same 
manner. 

Further  investigations  have  tended  to  diminish  the  impor- 
tance of  the  "mechanical"  factor  and  to  develop  a  selective 
or  secretory  function  for  the  cells,  even  including  those  of  the 
glomeruli,  by  virtue  of  which  the  cells  will  permit  certain 
substances  to  pass  through  and  prevent  others,  among  the 
latter,  albumin.  The  view  is  advanced  that  the  passage  of 
the  fluid  through  the  glomerulus  is  not  a  mere  transudation, 
but  a  matter  of  selection  also. 

The  selective  action  of  the  renal  cells  is  both  qualitative 
and  quantitative.  As  an  example  of  the  former,  it  is  shown 
by  experiment  that  if  some  egg  albumin  be  injected  into  the 
blood,  it  is  promptly  eliminated  by  the  kidneys.  Egg  albumin 
is  not  very  markedly  different  from  the  serum  albumin  of  the 
blood ;  both  are  indiffusible,  but  the  renal  cells  recognize  the 
former  as  a  foreign  substance  and  immediately  separate  it 
from  the  blood.  The  sugar,  in  normal  quantity  in  the  blood, 
although  a  diffusible  substance  is  not  selected.  Urea,  which 
is  also  diffusible,  but  existing  in  the  blood  in  much  smaller 
amount  than  sugar,  is  selected  and  appears  in  the  urine.  Why 
this  should  be  so  it  is  difficult  to  explain,  although  it  is  a  well- 
known  fact  that  the  sugar  serves  as  a  food  for  the  tissues  and 
is  needed  by  the  system,  while  urea  is  a  waste  product  and 
would  be  detrimental  to  the  system  if  not  eliminated. 

That  the  cells  exert  a  quantitative  selection  is  shown  by 
the  fact  that  when  sugar  is  present  in  the  blood  in  excessive 
amount,  3  parts  per  1000  or  over,  the  excess  is  promptly 
eliminated. 

Diet  influen(3es  the  reaction  of  the  urine.  A  vegetable 
diet  favors  an  alkaline  reaction ;  a  diet  of  flesh  favors  an  acid 
reaction.  The  urine  of  the  herbivora  is  therefore  alkaline, 
while  in  the  carnivora  and  omnivora  the  urine  is  mainly  acid, 
although  influenced  to  some  extent  by  the  kind  of  food  eaten. 


12 

The  formation  of  an  acid  fluid  from  alkaline  material, — blood 
and  lymph, — is  at  first  sight  puzzling.  The  well-known  fact 
that  the  gastric  secretion  of  all  mammals  is  acid  is  a  case  in 
point.  Furthermore,  experimental  evidence  shows  that  if  an 
alkaline  solution  of  sodium  bicarbonate  be  placed  upon  one 
side  of  the  membrane  in  an  osmometer  and  a  solution  of  neu- 
tral sodium  phosphate  be  placed  upon  the  other  side  and  a 
weak  electric  current  be  sent  through  the  solutions,  the  fluid 
in  contact  with  the  positive  pole  will,  in  a  short  time,  become 
acid  from  the  formation  of  acid  sodium  phosphate,  while  the 
fluid  in  contact  with  the  negative  pole  is  increased  in  alkalinity 
Na  HCOg+Na^  HPO,  =  Na,  COa-f  Na  H^  PO,. 

While  it  may  be  true  to  a  considerable  extent  that  the  kid- 
ney merely  removes  certain  constituents  from  the  blood  and 
transfers  them  to  the  urine,  it  has  been  shown  that  the  activity 
of  the  renal  cells  is  required  in  the  production  of  hippuric  acid, 
— a  new  product,  as  hippuric  acid  is  not  present  in  the  blood. 

In  general,  the  theory  explaining  the  secretion  of  urine, 
according  to  observed  facts,  is  one  which,  while  recognizing 
the  process  as  partly  physical,  also  requires  some  process  of 
activity  or  elaboration  on  the  part  of  the  kidney  itself. 


II. 

The  urine  of  all  mammals  may  be  regarded,  for  the  most 
part,  as  a  solution  of  constituents  derived  from  the  metabolism 
of  the  tissues  of  the  body.  Some  of  these  constituents,  espec- 
ially the  inorganic,  may  appear  in  the  urine  in  the  same  form 
as  they  are  taken  into  the  body  in  the  food,  e.  g.,  sodium 
chloride;  others,  especially  the  organic,  represent  decom- 
position products  derived  from  the  food  or  tissues,  e.  g.,  urea, 
creatinin,  etc.  The  composition  of  the  urine  may,  therefore, 
to  some  extent  be  regarded  as  an  index  of  tissue  activity,  and 
the  examination  of  this  secretion  is  of  considerable  importance 
in  clinical  diagnosis  and  prognosis. 

That  there  is  a  relationship  between  the  diet  and  the  renal 
excretion  is  shown  by  the  examination  of  the  urine  of  the  three 
great  classes  of  animals  grouped  according  to  the  food  they 
eat ;  herbivora,  omnivora  and  carnivora.    Perhaps  the  first  and 


13 

most  striking  characteristic  of  the  urine  of  each  group  is  its 
chemical  reaction.  In  vegetable  feeders  it  is  normally  alkaline ; 
in  flesh  eaters  it  is  markedly  acid,  and  in  the  omnivora  it  may 
be  acid  or  alkaline  according  to  the  preponderance  of  the 
fleshy  or  vegetable  material  in  the  food. 

In  the  practical  treatment  of  this  subject  it  is  convenient 
to  regard  the  horse  as  the  type  for  the  herbivora;  man  for 
the  omnivora ;  and  the  dog  or  the  cat  for  the  carnivora.  In  all 
cases  throughout  this  work  it  is  to  be  understood  that,  unless 
especially  directed,  the  same  tests  are  to  be  performed  upon 
the  urines  of  man  and  horse  and  a  parallel  record  of  such  tests, 
for  comparison,  is  to  be  kept  upon  the  blank  pages. 

In  man,  fresh,  normal  urine  is  a  clear,  golden-colored, 
transparent  liquid,  having  a  peculiar  aromatic  characteristic- 
odor,  and  a  bitter  saline  taste. 

In  the  horse,  the  urine  is  of  a  yellowish  color  when  passed, 
but  turns  to  a  deep  brown  color  upon  standing  for  a  time,  due 
to  the  oxidation  of  pyrocatechin.  It  is  more  or  less  turbid  and 
of  a  viscous  character.  Its  odor  is  somewhat  ammoniacal  and 
strongly  aromatic  and  more  penetrating  that  that  of  man. 

The  urine  is  chiefly  a  solution  of  urea  and  certain  organic 
and  inorganic  salts ;  epithelial  cells  and  mucus  may  also  be 
held  in  suspension.  Like  milk  and  other  animal  fluids,  the 
urine  is  not  of  constant  composition.  It  is  influenced  by  vari- 
ous factors,  such  as  food,  the  amount  of  water  or  other  fluids 
taken  into  the  body ;  the  temperature  of  the  skin ;  the  emo- 
tions ;  blood  pressure,  general  or  local ;  exercise ;  the  time  of 
day ;  age  ;  sex ;  and  medicines. 

Quantity.  The  amount  varies  considerably.  In  man.  the 
quantity  for  twenty-four  hours  ranges  from  1000  cc.  to  2000  cc. 
In  the  horse,  the  average  amount  is  about  3000  cc.  to  4000  cc, 
although  it  may  go  as  high  as  7000  cc.  or  9000  cc.  In  the  ox  a 
still  greater  quantity  is  secreted,  the  usual  limits  being  from 
4500  cc.  to  19000  cc.  In  the  sheep,  from  250  cc.  to  700  cc.  In 
the  pig  it  varies  from  1200  cc.  to  6000  cc.  In  the  dog  it  varies 
from  200  cc.  to  900  cc,  depending  upon  the  size  of  the  animal. 

Color.  The  color  ranges  from  pale  yellow  to  brown.  The 
normal  color  of  urine  is  due  to  pigments  probably  derived  from 
the  coloring  matter  of  the  bile. 


"  ■  tr  f  11^ 


14 

Transparency.  The  urine  of  the  horse  is  normally  more 
or  less  opaque,  that  of  man  should  be  transparent  at  the  time 
of  passing.  Many  pathological  urines,  however,  are  ;^erfectly 
clear.  Pathological  turbidity  may  be  due  to  urates,  phosphates, 
pus,  bacteria,  spermatozoa,  fatty  globules,  blood,  etV  All 
urines  become  turbid  after  standing  for  a  time.  \ 

The  urine  of  the  horse  is  especially  turbid  at  the  en^  of 
micturition,  exceptionally  it  may  be  clear  but  becomes  turbid 
on  cooling.  Its  opacity  is  due  to  the  presence  of  earthy  salfs 
of  the  carbonate  of  lime  precipitated  and  formed  by  the  disv 
engagement  of  a  certain  amount  of  carbon  dioxide  from  the 
bicarbonate  of  lime.  The  turbidity  increases  when  the  urine 
remains  for  any  length  of  time  in  the  bladder;  it  reaches  its 
maximum  when  the  urine  is  cooled  by  exposure  to  the  free  air  ; 
it  diminishes  after  the  ingestion  of  a  large  quantity  of  water. 

A  clear,  limpid  urine  is  generally  pathological  in  the  horse  ; 
it  indicates  polyuria  and  the  reaction  in  this  case  is  usually 
acid,  exceptionally  neutral  or  alkaline  when  the  phosphates  are 
modified  qualitatively  or  quantitatively. 

The  turbidity  of  horse  urine  is  abnormal  when  it  is  due 
to  the  presence  of  the  phosphate  of  lime,  or  of  calcium  sul- 
phate, or  acid  salts ;  also  from  the  existence  of  albuminoid  sub- 
stances, (exudates,  leucocytes  in  interstitial  nephritis). 

In  other  animals  the  passing  of  turbid  urine  is  usually  re- 
garded as  abnormal. 

Consistency.  In  the  horse  the  urine  is  very  viscid  on 
account  of  the  contained  mucus  and  sometimes  of  epithelial 
debris.  It  filters  very  slowly.  Urine  taken  directly  from  the 
ureter  or  the  pelvis  of  the  kidney  is  still  more  viscid,  having 
a  consistency  very  similar  to  that  of  egg  albumin  and  a  spe- 
cific gravity  somewhat  higher  than  normal. 

Reaction.  The  reaction  of  human  urine  is  acid,  that  of  the 
dog  more  so  than  that  of  man.  In  the  pig  it  is  sometimes  acid 
and  sometimes  alkaline,  depending  upon  the  diet.  In  the  horse 
and  sheep  it  is  alkaline,  also  in  the  ox,  but  in  the  calf  and  foal 
it  is  acid.  Herbivorous  urine  is  alkaline,  but  if  such  animals 
are  starved  for  a  time  they  practically  become  carnivorous  in 
that  they  are  living  upon  their  own  tissues  and  under  such  con- 
ditions their  urine  becomes  acid. 


15 

In  the  herbivora  the  reaction  is  normally  alkaline  and  is 
due  to  the  presence  of  bicarbonates  or  carbonates  of  lime  or 
potassium.  It  would  appear  that  the  salts  present  in  the 
vegetable  food  undergo  oxidation  to  form  organic  acids,  and 
these  in  turn  are  transformed  into  bicarbonates  or  carbonates, 
causing  alkalinity  of  the  urine. 

In  man  the  acidity  is  due  to  the  presence  of  acid  sodium 
phosphate  NaH^  PO^.  The  urine  passed  before  breakfast  and 
during  fasting  or  perspiration  is  more  acid  than  at  other  times ; 
during  digestion  and  after  meals  the  acidity  is  decreased. 

The  degree  of  acidity  of  the  urine  may  be  determined  by 
the  use  of  the  acidimeter.  a  graduated  glass  tube  devised  by  Dr. 
H.  R.  narrower.    His  description  of  it  and  its  use  follows: 

The  acidimeter  consists  of  a  glass  tube  so  graduated  that 
10  cc.  is  the  first  measuring  point.  From  this  upward  the  tube 
is  graduated  in  fifths  of  degrees  up  to  100°,  each  degree  repre- 
senting the  amount  of  decinormal  sodium  hydroxide  solution 
required  to  neutralize  100  ee.  of  urine.  The  method  of  using 
the  acidimeter  is  as  follows:  The  tube  is  filled  with  the  urine  to 
be  tested,  until  the  lower  edge  of  the  meniscus  is  just  on  the 
10  cc.  mark.  Two  drops  of  phenolphthalein  indicator  solution 
are  added,  and  then  with  an  ordinary  medicine  dropper  deci- 
normal sodium  hydroxide  solution  is  slowly  added,  inverting 
the  tube  after  each  addition,  until  the  color  of  the  fluid  has  just 
been  changed  from  a  yelloAv  to  a  light  rose  pink.  The  acidity  in 
degrees  is  now  read  off  on  the  tube  at  the  level  of  the  fluid.  The 
normal  acidity  of  a  mixed  21:  hour  specimen  should  be  between 
30  and  40  degrees. 

(With  very  concentrated  urines  in  which  the  acidity  is 
above  100°  the  tube  may  be  filled  to  the  5  cc.  mark  and  water  to 
the  usual  level.    The  resulting  figures  are,  of  course,  doubled.) 

If  the  urine  is  alkaline  in  reaction  and  it  is  desired  to  esti- 
mate the  degree  of  alkalinity  decinormal  hydrochloric  or  oxalic 
acid  solution  must  be  used  in  place  of  the  sodium  hydroxide, 
the  pink  color  present  being  just  discharged  by  the  acid." 

The  "Acid  Index"  or  "Acid  Unit"  may  be  obtained  by 
multiplying  the  degree  of  acidity  by  the  amount  of  urine  passed 
in  24  hours.    The  normal  in  man  is  about  40,000  acid  units. 

In  man  there  is  increased  acidity  physiologically  during  the 


16 

night ;  with  a  flesh  diet ;  after  strong  muscular  exertion ;  during^ 
the  intervals  of  gastric  digestion ;  after  the  ingestion  of  mineral 
acids. 

There  is  increased  acidity,  pathologically,  in  fevers ;  in  rheu- 
matism ;  after  asthmatic  attacks ;  in  emphysema,  pneumonia 
and  pleuritis. 

The  urine  is  less  acid,  or  alkaline,  physiologically,  during 
gastric  digestion ;  after  hot  or  prolonged  cold  baths ;  after  pro- 
fuse sweating;  after  copious  ingestion  of  vegetable  acids  and 
their  salts. 

Pathologically  in  acute  and  chronic  inflammation  of  the- 
urinary  tract  as  in  cystitis ;  in  decomposition  of  the  urine  in  the 
bladder  in  retention ;  in  some  cerebral  and  nervous  diseases ;  in 
anemia ;  chlorosis ;  and  general  debility. 

When  the  urine  of  man  is  set  aside  in  a  cool  place  it  grad- 
ually becomes  more  acid.  This  is  called  acid  fermentation.. 
After  longer  exposure  to  a  warm  atmosphere  the  urine  becomes 
neutral,  and  finally  strongly  alkaline  in  reaction.  It  becomes 
turbid,  has  an  ammoniacal  odor,  and  deposits  triple  phosphate, 
ammonium  urate,  and  great  numbers  of  microbes  exist.  This  is 
alkaline  fermentation  and  is  due  to  the  transformation  of  the 
urea  into  ammonia  and  carbon  dioxide,  by  means  of  a  ferment 
produced  by  an  organism  known  as  the  micrococcus  ureae.  (Fig. 
20).  This  organism  is  said  to  be  conveyed  through  the  air  and 
to  exist  commonly  around  the  orifice  of  the  urethra.  As  long 
as  the  urine  is  acid  the  organism  does  not  exist  in  the  bladder, 
but  may  sometimes  gain  entrance  through  the  medium  of  a 
sound  or  catheter. 

Test  the  reaction  of  the    urines  with    red    and    blue 

litmus  paper.     Some  urines   change    both    the    red    and 

blue  paper  and  are  termed  amphoteric.     (In  taking  notes 

of  the  experiments    it    is  well    to    record   the    results    in 

parallel  columns,  the  horse  urine  in  one  column  and    the 

human  urine  in  the  other).  ^ 

Specific  Gravity.     The    specific    gravity  of    human    urine 

ranges  from  1015  to  1025,  the  average  being  1018  to  1020.  That 

of  the  horse  ranges  from  1020  to  1050,  the  average  being  about 

1035.    That  of  the  cow  is  lower,  ranging  from  1015  to  1045.    It 

seems  to  depend  considerably  upon  the  milk  secreted.    In  milch 


\r^ 


17 


C3WS  the  urine  contains  a  greater  amount  of  water  and  a  lesser 
amount  of  solids.  The  specific  gravity  of  the  urine  of  the  sheep 
ranges  from  1015  to  1060 ;  that  of  the  pig  from  1005  to  1025,  and 
of  the  dog  from  1016  to  1060,  depending  upon  the  diet.  Cat, 
1020  to  1040. 

The  specific  gravity  may  be  ob- 
tained in  different  ways.  The  sim- 
plest and  most  usual  way  is  to  employ 
the  instrument  known  as  the  urin- 
ometer.  Some  urinometers  are  not 
strictly  accurate,  but  they  may  be 
tested  by  filling  the  urinometer  jar 
with  distilled  water  at  15°  C.  (60°  F.) 
Read  the  division  of  the  scale  corre- 
sponding with  the  surface  of  the 
fluid  looking  above  or  below  the 
meniscus  as  is  found  to  be  the  most 
correct  for  the  zero  reading.  Always 
adhere  to  this  method  when  using 
the  same  urinometer.  Test  the  spe- 
cific gravity  of  the  urines  and  make 
the  necessary  corrections.  If  the 
urine  is  warmer  than  15°  C.  add  1  to 
the  last  right  hand  figure  of  the  spe- 
cific gravity  for  every  4  degrees  of  C. 
temperature,  or  for  every  7  degrees 
__  of  extra  F.  temperature.    As  oppor- 

tunity presents,  test  the  specific  grav- 
ity of  some  warm,  freshly  passed 
urine.  Test  the  same  urine  later, 
when  cool,  and  note  if  any  difference 
in  the  reading. 

Urinometers  already  corrected  for  the  ordinary  room  tem- 
perature (70°  F.)  may  be  obtained,  in  which  case  the  temper- 
ature corrections  may  be  omitted.  If  the  urine  should  be  too 
dense  to  read  easily  on  the  urinometer,  dilute  it  with  an  equal 
volume  of  distilled  water  and  multiply  the  reading  by  two  to 
get  the  correct  specific  gravity.  The  variation  of  the  specific 
gravity  depends  upon  the  amount  of  the  solids  in  the  urine. 
The  amount  of  solids    may  be    estimated    with    approximate 


18 

accuracy  from  the  specific  gravity  by  Christison's  formula 
(Haser-Trapp's  coefficient)  :  "Multiply  the  last  two  figures  of 
a  specific  gravity  expressed  in  four  figures  by  2.33.  This  gives 
the  quantity  of  solid  matter  in  every  1000  parts."  (The  num- 
ber of  grams  in  1000  cc). 

Example.  Suppose  a  patient  passes  1400  cc.  of  urine  in 
24  hours  and  the  specific  gravity  is  1020,  20X2.33  =  46.6  grams 
of  solids  in  1000  cc.  To  ascertain  the  amount  in  1400  cc.  use 
the  following  proportion :  1000  cc.  :  1400  cc.  : :  46.6  grams  is 
to  X  (65.24  grams).  The  total  quantity  of  the  solids  of  the 
human  urine  is  about  60  grams  for  the  24  hours  or  approxi- 
mately 4%. 

The  following  method  taken  from  the  Alkaloidal  Clinic  may 
also  be  used :  Multiply  the  quantity,  in  ounces,  of  the  24-hour 
urine  by  the  last  two  figures  of  the  specific  gravity  and  this 
by  1.1,  the  product  will  represent  the  total  solids  in  grains. 
Thus,  if  the  amount  of  urine  voided  in  24  hours  be  36  ounces 
and  its  specific  gravity  1021,  the  formula  would  be  36X21X1-1, 
equal  to  831  grains,  the  normal  amount  for  a  person  weighing 
100  lbs.  The  amount  for  other  weights  may  be  determined  by 
proportion. 

In  general  the  amount  of  total  solids  is  a  measure  (a)  of 
the  activity  of  tissue  change;  (b)  of  renal  integrity;  (c)  of 
abnormal  constituents  in  the  urine.  Hygienic  conditions  which 
favor  increased  metabolism,  as  abundant  food,  active  exercise, 
etc.,  increase  the  solid  matter  in  urine,  while  the  opposite  con- 
ditions decrease  them. 

With  the  urine  normal  or  subnormal  in  amount,  the  solids 
are  deficient  pathologically  from  defective  and  enfeeMed  meta- 
bolism as  in  senility ;  anemia  as  a  result  of  syphilis,  cancer,  etc. ; 
chronic  alcoholism;  functional  or  organic  diseases  of  the  liver. 
From  renal  failure  as  in  acute  nephritis ;  certain  conditions 
of  chronic  renal  disease ;  at  the  close  of  Bright 's  disease ; 
venous  congestion  of  the  kidneys,  etc.  With  the  urine  in- 
creased in  amount  there  may  be  a  deficiency  of  solids  in  dia- 
betes insipidus;  interstitial  nephritis;  amyloid  disease  of  the 
kidney;  chronic  parenchymatous  nephritis.  When  the  urine 
is  not  increased  in  amount,  the  urinary  solids  are  increased  in 
fevers;  lithemia;  some  forms  of  dyspepsia.     When  the    quan- 


'nruy   r^    vol    ^  >K-«— 


19 

tity  of  urine  is  increased  the  solids  are  increased  in  diabetes 
mellitus;  phosphaturia ;  azotiiria  (excessive  secretion  of  urea). 
Calculate  the  solids  of  the  urines  by  the  formulae 
previously  given.  After  obtaining  the  result  for  1000  cc. 
estimate  the  quantity  in  1250  cc.  of  human,  and  5450  cc. 
of  horse  urine.  In  recording  the  tests  use  parallel  col- 
umns, one  column  for  the  horse  and  the  other  for  the  hu- 
man urine. 


III. 

Qualitative  Tests.     In  all  cases  the  urines  must  be  filtered 
and  perfectly  clear  before  attempting  the  examination. 


INORGANIC  CONSTITUENTS. 

These  consist  chiefly  of  sodium,  potassium,  ammonium,, 
calcium,  and  magnesium,  combined  with  hydrochloric,  phos- 
phoric and  sulphuric  acids. 

Water.  The  water  of  the  urine  is  derived  from  the  food 
and  drink,  a  small  quantity  being  formed  in  the  body.  It  var- 
ies according  to  the  activity  of  the  sweat  glands  of  the   skin. 

Chlorides.  Next  to  the  urea  the  chlorides  form  the  chief 
portion  of  the  urinary  solids.  The  chlorides  are  increased 
physiologically  after  the  ingestion  of  salt  foods  and  much 
water ;  mental  and  physical  activities ;  and  during  pregnancy. 
Pathologically,  they  may  increase  after  the  crises  of  fevers ; 
after  the  absorption  of  exudates;  in  diabetes   (occasionally). 

The  chlorides  are  decreased  pathologically,  in  all  acute 
fevers;  pneumonia  (often  entirely  absent  during  the  height 
of  the  disease)  ;  in  cholera;  and  in  most  chronic  diseases.  An 
increase,  or  the  re-establishment  of  the  excretion  of  chlorides 
in  disease  is  generally  a  favorable  sign.  In  pneumonia  it  is 
a  precursor  of  the  crisis,  and  may  often  take  place  before  other 
symptoms  reveal  the  favorable  change. 

Test  a  portion  of  each  urine  with  a  few  drops  of 
silver  nitrate  solution.  A  white,  cheesy  or  curdy  pre- 
cipitate insoluble  in   nitric    acid   indicates  the  presence 


20 

of  silver  chloride.  The  phosphate  of  silver  may  also 
be  throM^n  down  but  the  nitric  acid  dissolves  it,  keeping 
it  in  solution. 

Evaporate   carefully   a  few  drops   of  urine  upon   a 
glass  slide,  with  a  gentle  heat  over  the    flame.     Octa- 
hedral or  rhombic   crystals  may  form,— a  compound  of 
sodium  chloride  and    urea.     Examine  with    the    micro- 
scope.    (Fig.  5). 
Sulphates.     The  sulphates  are    chiefly    those    of    sodium 
and  potassium.    Only  a  small  amount  of  them  enters  the  body 
with  the  food,  so  that  they  are  chiefly  formed  from  the  meta- 
bolism of    proteids  in  the  body.     The    above    are    known    as 
ordinary  sulphates.    Another  class  known  as  the  ethereal  sul- 
phates also  exist.     The  proportion  exists  in  the   ratio   of   10  of 
the  ordinary  to  1  of  the  ethereal  in  man.     In    the   horse   the 
proportion  is  about  2  of  the  ethereal  to  1  of  the  ordinary.    The 
ethereal  sulphates  are  formed  by  the  combination  of  sulphuric 
acid  with  organic  bases  such    as    phenol,  skatol,  etc.,  which 
originate  from  putrefactive  processes  in  the    intestine.     The 
amount  of  ethereal  sulphates  is  of  importance  in  determining 
whether  or  not  the  digestive  processes  are  going  on  normally. 
In  general  the  sulphates  are  increased  physiologically  by  the 
ingestion  of  sulphur  and  its  compounds ;  nitrogenous  food ;  and 
•conditions  of  increased  metabolism. 

After  acidulating  the  urine  with  hydrochloric  acid 
to  prevent  the  precipitation  of  phosphates,  add  to  a  small 
part  of  each  urine,  a  little  2%  barium  chloride  solution; 
a  precipitate  of  barium  sulphate  is  formed,  insoluble  in 
nitric  acid. 

To  separate  the  ethereal  sulphates,  mix  30  cc.  of 
urine  with  an  equal  bulk  of  "baryta"  mixture.  Stir  and 
filter.  This  removes  the  ordinary  sulphates  (as  barium 
sulphate),  add  10  cc.  of  hydrochloric  acid  to  the  above 
filtrate,  and  keep  in  the  water  bath  at  100°  C.  for  an 
hour  in  the  hood  and  then  allow  the  ethereal  sulphates 
to  settle.  This  may  require  some  little  time.  (Baryta 
mixture  is  prepared  by  making  saturated  solutions  in 
the  cold,  of  barium  nitrate  and  barium  hydrate,  and 
adding  two  volumes  of  the  hydrate  to  one  volume  of  the 
nitrate). 


'^K  vj;Ap\    v^i^  nA«^  :wVt:;  o\ 


21 

Phosphates.  The  phosphates  consist  of  alkaline  and 
earthy  salts  in  the  proportion  of  2  to  1.  The  latter  are  insol- 
uble in  an  alkaline  medium  and  are  precipitated  when  acid 
urine  becomes  alkaline.  They  are  insoluble  in  water,  but 
soluble  in  acids ;  in  acid  urine  they  are  held  in  solution  by  free 
CO2.  The  alkaline  phosphates  (sodium  and  potassium)  are 
very  soluble  in  water,  and  they  never  foriii  urinary  deposits. 
The  earthy  are  phosphates  of  calcium  (Cag  P0^)o  (abundant) 
and  magnesium  (MgHPO^  plus  THaO)  (scanty).  An  alkaline 
medium  precipitates  them  although  not  in  the  form  in  which 
they  occur  in  the  urine. 

The  excretion  of  phosphates  in  the  urine  is  largely  de- 
pendent upon  the  amount  of  calcium  ingested;  the  more  cal- 
cium the  food  contains,  the  less  phosphoric  acid  appears  in  the 
urine,  and  the  more  in  the  feces.  This  is  due  on  the  one  hand 
to  the  tendency  on  the  part  of  calcium  to  form  insoluble  cal- 
cium phosphates  in  the  intestinal  tract  and  in  this  way  to 
prevent  the  absorption  of  the  food  phosphates ;  on  the  other 
hand,  to  the  well-established  tendency  6f  calcium  salts  to  be 
excreted  into  the  bowel  and  not  into  the  bladder;  one  must 
imagine  in  the  latter  case  that  calcium  salts  circulating  in  the 
blood  combine  with  circulating  phosphoric  acid  and  bear  the 
latter  with  them  into  the  bowel. 

The  fact  is  of  some  therapeutic  importance  in  the  treat- 
ment of  nephrolithiasis  due  to  uric  acid  calculi,  for  the  admin- 
istration of  calcium  salts  in  this  affection  by  bearing  much 
phosphoric  acid  into  the  bowel,  leads  to  the  excretion  of  less 
phosphoric  acid  in  the  urine,  and  hence  of  normal  and  basic 
instead  of  acid  phosphates;  and  as  the  latter  precipitate  and 
the  former  dissolve  uric  acid,  it  will  be  spen  that  by  giving 
calcium  we  prevent  the  precipitatipn  of  crystalline  uric  acid 
and  urate  deposits  in  the  urinary  passages.     (Croftan). 

"Where  considerable  calcium  is  present  in  the  food  the 
excretion  of  phosphates  in  the  urine  is  minimum,  especially  if 
the  urine  is  alkaline  because  of  the  presence  of  sodium  or 
potassium  salts.  In  herbivorous  animals  where  the  urine  is 
alkaline  and  where  considerable  quantities  of  phosphorus  con- 
taining food  are  eaten,  very  little  phosphate  is  excreted  in  the 
urine. 


22 

Phosphates  are  increased  in  the  urine  pathologically  in 
rickets;  osteomalacia;  osteoporosis;  fractures;  chronic  rheu- 
matism; diseases  of  the  nervous  system;  and  after  great 
mental  strain  and  worry.  Phosphates  are  decreased  in  renal 
diseases  and  phthisis. 

Physiologically,  variations  occur  chiefly  from  the  character 
of  the  food  and  drink ;  in  the  horse  there  may  be  an   increase 
in  the  urinary  phosphates  after  a  large  feed  of  oats,  bran,  oil- 
cake, etc.    Pathologically,  they  are  increased  during  the  active 
changes  in  such  bone  diseases  as  spavin,  ring-bone,  splint,  etc. 
To  a  small  amount  of  each  urine  add  about  half  its 
volume  of  nitric  acid  and  then  add  two  volumes  of  a  5% 
solution  of  ammonium  molybdate  and  boil.     A    canary 
yellow  ppt.   (crystalline)  of  ammonium  phospho-molyb- 
date  should  appear    in  the    omnivorous  urine.     In    the 
herbivorous  urine  the  presence  of  so  much  organic  mat- 
ter renders  the  test  unreliable;  although  a  precipitate 
may  form  it  is  not  a  typical  or  characteristic    one    for 
phosphates  in  the  urine  of  the  horse,  unless  the  organic 
matter  has  been  previously  removed. 

To  each  of  the  urines  add  half  its  volume  of  am- 
monia and  allow  it  to  stand.  A  precipitate  of  earthy 
phosphates  is  formed,  in  the  urine  of  man.  Filter,  add 
enough  nitric  acid  to  give  an  acid  reaction  to  the  urines, 
and  test  the  filtrates  with  ammonium  molybdate  as 
before.  This  method  separates  the  earthy  from  the 
alkaline  phosphates. 

To  a  portion  of  the  urines  add  half  their  volumes 
of  baryta  mixture;  a  copious  precipitate.  Filter,  add 
nitric  acid  and  test  the  filtrates  with  ammonium  molyb- 
date. No  ppt.  should  occur  as  the  baryta  mixture  pre- 
cipitates the  phosphates  as  well  as  the  sulphates  and 
carbonates. 

Use  a  little  of  the  magnesia  mixture  instead  of  the 
baryta  mixture.  Filter,  add  a  little  nitric  acid,  and  test 
the  filtrate  with  ammonium  molybdate.  (The  magnesia 
mixture  is  composed  of  magnesium  sulphate  1  part, 
ammonium  chloride  1  part,  ammonia  water  1  part,  and 
distilled  water  8  parts). 


w.^ 


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^f^WtM^"^  I^V^l      Ovv  .VC S.  \A\C^1^^V^.> 


vv\3. 


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ffy^    '>AAN\5. 


23 

To  portions  of  the  urines  add  a  few  drops  of  acetic 
acid  and  then  a  little  5%  uranium  nitrate  solution, — a 
yellow  ppt.  of  uranium  phosphate  is  formed. 
The    lime,  magnesia,  iron    and    other    inorganic    urinary 
constituents  are  comparatively  unimportant,  and  have  no  spe- 
cial clinical  significance.     The  tests  for  them  are  somewhat 
complicated  and  are  therefore  omitted. 

Demonstration  of  Carbonates  and  COo  in  Urine.  Carbon 
dioxide  exists  in  the  urine  to  some  extent  in  a  free  state.  There 
are  also  various  carbonates  present,  especially  in  the  herbivor- 
ous urine.  The  amount  is  very  variable  and  to  a  great  extent  is 
dependent  on  the  kind  of  food  that  is  eaten ;  large  quantities  of 
vegetable  foods  determine  an  increase  both  in  the  combined  and 
in  the  free  carbon  dioxide.  The  carbonates  may  be  broken  up 
.and  COo  given  off  by  the  application  of  heat  or  certain  acids. 

Heat  experiment.  Fill  a  small  flask  about  half  full 
of  unfiltered  herbivorous  urine.  Through  the  perforated 
stopper  of  the  flask  pass  some  bent  glass  tubing  con- 
nected with  a  test  tube  containing  lime  water.  Heat  the 
urine  in  the  flask,  and  as  it  boils  the  CO,  will  pass  over 
into  the  test  tube,  and  calcium  carbonate  will  be  formed 
from  the  union  of  the  gas  with  the  lime.  Repeat  the  ex- 
periment with  omnivorous  urine. 

A  simple  proof  of  the  presence  of  carbonates,  or 
CO2  jji  the  urine  is  to  fill  a  Doremus  Ureometer  with 
25  ce.  of  the  unfiltered  urine  and  introduce  1  cc.  of  nitric 
acid.  The  larger  part  of  the  gas  (CO,)  rises  and  col- 
lects in  the  upper  portion  of  the  ureometer.  Compare 
the  amounts  thus  obtained  in  the  urines  of  horse  and 
man. 

A  simpler  qualitative  test  is  to  add  a  few  drops  of 
nitric  or  acetic  acid  to  a  little  unfiltered  urine  in  a  test 
tube.  If  effervescence  occurs  it  is  due  to  CO2  set  free 
from  the  carbonates  by  the  acid.  '• 


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24 

IV. 

ORGANIC  CONSTITUENTS. 

Urea  is,  in  amount,  the  principal  constituent  of  the  solids, 
of  the  urine.  It  is  the  most  important  product  of  the  decomposi- 
tion of  proteid  in  the  food.  In  round  numbers  it  forms  from 
2%  to  3.5%  of  the  urine,  averaging  about  2.5%.  About  one-half 
of  the  total  solids  in  the  urine  consists  of  urea.  Urea  has  no  effect 
on  litmus,  it  is  odorless,  has  a  weakly  cool  and  bitter  taste  like 
saltpeter.  It  is  verj^  soluble  in  water  and  alcohol,  but  it  is  almost 
insoluble  in  ether  and  benzine.  About  90%  of  the  total  nitrogen 
of  the  urine  is  excreted  in  the  form  of  urea. 

Physiologically,  urea  is  increased  by  a  proteid  diet ; -exercise 
and  muscular  vigor;  by  drinking  much  water.  It  is  decreased  by 
fasting;  noq-nitrogenous  food;  reduction  of  water  in  the  d(et ; 
alcoholic  beverages,  tea  or  coffee;  indolence  of  mind  and  boidy. 
Pathologically,  it  is  increased  in  all  acute  fevers;  dyspnoea; 
diabetes;  and  phosphorus  poisoning.  It  is  decreased  in, uremia; 
acute  yellow  atrophy  of  the  liver  and  in  chronic  diseases.  In 
general,  any  disease  that  interferes  with  the  activity  of  the  liver 
.decreases  the  urea.  Any  disease  affecting  the  uriniferous  tubules 
"  may  modify  the  appropriation  of  the  urea  from  the  blood  and 
affect  its  passag:e  into  the  urine.  An  increase  of  the  urea  inde- 
pendently of  the  physiologic  variations  and  aniount  of  nitro- 
genous food  eaten  is  an  approximate  index  of  the  amount  of 
tissue  waste  in  the  system;  on  the  other  hand,  when  the  urea  is 
decreased  it  is  an  evidence  of  a  diseased  condition  of  the  liver 
(the  producer  of  urea)  or  the  kidney  (the  eliminator  of  urea). 

A  simple  method  of  detecting  urea  is  to  concentrate 
a  small  amount  of  dog  *  or  human  urine  in  an  evaporating 
dish  to  about  half  of  its  .original  volume.  Place  a  drop 
or  two  of  this  concentrated  urine  upon  a  glass  slide  and 
■  after  adding  a  drop  of  nitric  acid,  gently  warm  over  the 
flame.  If  urea  be  present,  upon  evaporation,  the  micro- 
scope will  show  the  characteristic  crystals  of  nitrate  of 
urea,  of  rhombic  or  hexagonal  form.    (Fig.  4). 


*  For  this  experiment,  dog  urine  is  more  satisfactory  than  humart 
because  of  the  larger  percentage  of  urea  present. 


\)W  ca^J3a   M^  U(U!c.till 


25 


Take  20  cc.  of  fresh,  filtered  dog  or  human  urine  and 
add  20  cc.  of  baryta  mixture  to  precipitate  the  phosphates 
and  sulphates.  Filter,  evaporate  the  filtrate  to  dryness, 
and  extract  the  residue  with  a  little  boiling  alcohol  over 
the  water  bath  very  carefully.  Filter  off  the  alcoholic 
solution,  place  some  of  it  on  a  slide  and  allow  the  crystals 
of  urea,  usually  long,  fine,  transparent  needles,  to  separate 
out.    Examine  them  under  the  microscope.     (Fig.  5). 

Repeat  in  the  hood,  the  experiment  upon  the  urine  of 
the  horse  and  compare  results. 

Heat  some  urea  crystals  in  a  test    tube.     Biuret    is 
formed  and  ammonia  comes  off.     Add  a  trace  of  copper 
sulphate  solution  and  a  few  drops  of  20%  caustic  potash. 
A  rose-red  color  is  produced, — the  biuret  reaction. 
Medicines  which  increase  the  amount    of  urea    are :    urea 
itself;  uric    acid,  common    salt,  phosphoric    acid,  squill,  theo- 
bromine,   colchicum,    cubebs,    atropine,     cantharides,    vegetable 
acids,  iron  preparations,  hyposulphite  of  soda,  potassium  chlor- 
ide, ammonium  chloride,  coca,  potassium  permanganate,  oxygen, 
salicylic  acid. 


Fig.  4. 
Crystals  of  Nitrate  of  Urea. 


Fig.  5. 
Crystals  of  Urea. 


Medicines  which  decrease  the  amount  of  urea  are :  digitalis, 
alcohol,  coffee,  tea,  potassium  and  sodium  iodides,  potassium 
bromide,  arsenic,  turpentine,  alkaline  carbonates,  mercury,  anti- 
pyrin,  valerian,  quinine  sulphate,  benzoic  acid. 


W  2mAS^  e4^  '^  '^-^"^ 


2«; 

Uric  Acid,  sometimes  called  litliic  acid,  is,  next  to  urea, 
the  most  important  nitrogenous  constituent  of  the  urine  of  man. 
It  has  been  said  to  be  absent  in  herbivorous  urine,  being  replaced 
by  hippuric.  This  has  been  shown  by  later  researches  not  to  be 
strictly  true,  as  a  trace  of  uric  acid  is  found  in  addition  to  the 
hippuric.  In  birds  and  reptiles  uric  acid  is  the  chief  nitrogenous 
constituent,  being  present  in  greater  amount  than  urea. 

In  man  the  proportion  of  urea  to  uric  acid  is  about  45  to 
^1,  ab^t  0.-5  gr^niaf  the  latter  being  excreted  in  i^e  24  hours. 

e^usei'.the  .OTi(5k  r^^. deposit- ■sx)meti'ines  §^n  -^n^'Tirine  after  ■: 
Standing  for  a  time.  Its  solubility.is  very  low,  only  1  part  bein^ 
V  'soluble  in.  14000  cc.  of.-fcdld  wat%  and  Ito  ^^^SQOQ'Vin  boiling.:  v.. 
Physiologically,  it  is  increased  and  diminished  pVoportionfrgl;^*' 
.  with  urea.  -Pathologically,  it, is^' increased  in^ indigestion;  acuteC>^ 
dropsies,  rheumatic  and' catarrhal  inflamrftcations ;  aftfer  attackg'-'  v'W4.- 
of  gout ;  cancer  of  the  liver ;  in  leucemia ;  in  a}l  disturbances  of 
the  circulation  and  respfiifatidn.'  Pathologically  uric  acid  is  de- 
creased in  chronic  diseases  generally ;  diabetes  and  polyuria ;  be- 
fore paroxysms  of  gout ;  anemias ;  chronic  rheumatism ;  chronic 
diseases  of  spinal  cord. 

Uric  acid  is  generally  in  solution  in  the  form  of  urates    of 
sodium,  potassium,  ammonium,  lime  and  magnesium.    These  salts 
are  very  easily  decomposed  even  by  weak  organic  acids. 
Perform  the  following  tests  upon  both  urines  (filtered). 

In  a  conical  glass,  add  5  parts  of  hydrochloric  acid 
to  30  parts  of  urine.  Label  and  put  in  a  cool  place  for 
24  hours.  Eed  or  brownish  colored  crystals  of  uric  acid 
are  deposited  upon  the  sides  of  the  glass,  or  form  a 
pellicle  on  the  surface  of  the  fluid  like  fine  grains  of 
cayenne  pepper.  The  brownish-red  color  is  due  to  pig- 
ment (uroerythrin). 

Murexide  Test.  To  about  1  cc.  of  human  urine  add 
a  little  nitric  acid;  evaporate  in  a  porcelain  dish  very 
carefully  to  avoid  charring.  Cool  and  add  a  drop  of 
ammonia,  a  purple  red  color  of  murexide  or  purpurate 
of  ammonia  is  formed.  It  turns  bluer  upon  the  addition 
of  caustic  potash  solution. 

Dissolve  a  few  crystals  of  uric  acid  in  10%  caustic 
soda  or  potash.     Add  a  drop  or  two  of  Fehling's  Solu- 


27 


tion — or  dilute  cupric  sulphate  and  caustic  potash  and 
heat — there  should  occur  a  ppt.  which  at  first  may  be 
white  and  after  a  time  turning  green  or  reddish. 

(Fehling's  Solution  is  put  up  in  two  bottles;  one 
labeled  A,  the  other  B.  In  making  tests,  take  equal  parts 
of  A  and  B  and  add  the  substance  to  be  tested.  See  for- 
mula in  appendix.) 

Schiff's  Test.     Dissolve  a  little  uric  acid  in  a  small 
quantity  of  10%  sodium  carbonate  solution.    "With  a  glass- 
rod,  place  a  drop  of  silver  nitrate  solution  on  filter  paper 
and  then  a  drop  of  the  uric  acid  solution  so  that  the  two 
drops  partially  overlap.     A  dark  brown  or  black  spot  of 
reducing  silver  appears. 
Hippuric    Acid,     (C9H9NO3),    is    found    especially  in    the 
urine  of  the  herbivora,  as  the  horse,  ox,  etc.     In   the   urine   of 
carnivora,  and  especially  in  that  of  man,  it  exists  in  but  very 
minute  quantity,  usually  abouV 0.5  to  1  gram  being  excrete^  in  ■'<■ 

2i  hours. -It  dissolves  readily  in  hot  alcoliQl  but  is  spajringly  s^-'    JtA  J 
uble  in  water.     It  occurs  in  man  after  the  ingestion  of  certain  ' 

vegetables,  such  as  asparagus,,  plums,  pears,  and  apples  with        ^         , 
their  skins;  a  ptirely  vegetable  diet  and  from  the  use  of  benzoic  "> 

acid,  cinnamic  acid,  essence  of  bitter  almonds,  quinine,  and  an- 
alogous bodies.  Hippuric  acid  normally 'seems  to  be  derived 
chiefly  from  the  husks  or  cuticular  structui^ps  of  the  food. 

Pathologically  hi-p-pnric  acid  isincreased  in  diabetes,  chorea ; 
jaundice  and  other  liver  complaints ;  and  in  the  acid  urine    of 


.A 


Fig;,  6. 
Crystals  of  Hippurio  Acid. 


Fig.  7. 
Various  forms  of  Hippuric  Acid 
with  triple  phosphates. 


I^^Suh^  xWS^  oJ^  \XyA^  ^/^  ;^SmM  vnA 


28 

patients  suffering  from  all  kinds^of  J^evers.  In  testing  for  hip- 
purie  acid  the  fresh  urine  shonld'l3e  used;  if  stale,  benzoic  acid  is 
likely  to  be  obtained  instead. 

Test.  Saturate  the  fresh  urine  with  lime  water,  which 
transforms  the'  hippuric  acid  into  a  salt  of  lime,  the  fluid 
is  then  filtered,  evaporated  to  a  syrupy  consistency  and 
excess  of  hydrochloric  acid  added,  when  hippuric  acid 
crystallizes  out  on  standing.  The  horse  urine  is  to  be 
evaporated  in  the  hood. 

Creatinin  (C4H7N3O).  This  substance  was  discovered  in 
the  urine  by  Liebig.  It  is  easily  produced  from  creatin,  a  sub- 
stance normally  existing  in  plain  and  striated  muscular  tissue. 
Creatinin  occurs  constantly  in  normal  human  urine,  the  amount 
varying  according  to  Voit,  from  0.5  to  4.9  grams  per  day  accord- 
ing to  the  quantity  of  proteids  eaten.  It  is  said  not  to  be  dimin- 
ished by  fasting. 

Pathologically  it  is  increased  in 
typhoid  fever;  intermittent  fever; 
pneumonia;  tetanus.  It  is  de- 
creased during  convalescence  from 
the  above  diseases ;  and  likewise 
in  anemia ;  chlorosis ;  muscular 
atrophy ;  tuberculosis ;  paralysis, 
etc. 

Test.  Add  a  little  caustic  soda 
to  the  urine  and  then  a  few  drops 
of  freshly  prepared  1%  sodium 
nitro-prusside.  A  ruby  red  color 
develops.  Boil,  the  color  fades.  "While  boiling,  add  glacial 
acetic  acid,  the  color  changes  to  blue.  The  above  is  a  modi- 
fication of  Weyl's  test.  Acetone,  if  present,  also  gives  the 
red  reaction. 

Perform  the  above  test  on  both  urines. 
Caution.     Creatinin  reduces  copper  oxide  and  may 
be  taken  for  small  quantities  of  sugar. 
Mucus.     Mucus    in    the    urine  is    not  visible,  but    causes 
cloudiness  sometimes  by  entangling  epithelial  cells,  urates,  oxalate 
of  lime  and  other  crystals  in  various  amounts. 

Add  to  the  urine  a  little  acetic  or  citric  acid  and,  in 


.v<^  ^n^^^XAm^  ^^  Moftu 


UA,  •  * 


29 

addition,  a  few  drops  of  liquor  iodi  comp.     (Lugol's  solu- 
tion) which  makes  the  threads  or  bands  of  mucin  visible. 

Indican  or  Indoxyl.  This  substance  is  derived  from  indol, 
one  of  the  putrefactive  products  formed  in  the  intestine.  Indoxyl 
occurs  in  very  small  quantities  in  normal  human  urine,  about 
.004  to  .020  gram  for  the  24  hours.  Horse's  urine  is  said  to  con- 
tain 23  times  as  much.  The  intestines  of  the  herbivora  are  much 
longer  than  in  the  ease  of  the  carnivora.  On  this  account,  and  in 
conjunction  with  the  carbohydrate  diet,  a  much  greater  fermen- 
tation occurs,  which  leads  to  a  greater  elimination  of  indican  in 
the  urine. 

In  obstruction  of  the  intestine,  or  in  intestinal  catarrh  or 
where  the  food  remains  a  long  time  in  the  intestine  and  ferments 
there,  the  proportion  of  indican  increases  in  the  urine  and  causes 
a  true  indicanuria.  Indoxyl  is  of  considerable  clinical  import- 
ance, an  increase  is  indicative  of  imperfect  performance  of  the 
digestive  processes.  In  obstructive  diseases  of  the  small  intestine 
the  increase  of  indoxyl  in  the  urine  is  enormous. 

Pathologically  indoxyl  is  increased  in  cholera,  typhoid  fever, 
peritonitis,  dysentery,  Addison's  disease,  cancer  of  the  liver  and 
stomach  and  pernicious  anemia.- 

^  Jaffe's  Test  ■  To  a.  little  urine  add  an  equal  volume 
of  strong  hydrochloric  acid.  Add  to  this  mixture  2  or  3 
drops  of  a  solution  of  freshly  prepared,  chlorinated  soda. 
There  soon  forms  a  bluish  cloud  of  indigo.  Add  a  little 
chloroform  and  shake  gently,  this  will  take  the  indigo  into 
solution  and  settle  as  a  blue  layer  at  the  bottom  of  the  test 
tube.  The  amount  of  indoxyl  can  be  judged  by  the  depth 
of  the  blue  color.  Indican  is  oxidized  by  free  chlorine  ob- 
tained from  the  chlorinated  soda  to  indigo. 

The  Hydrochloric  acid— Ferric  Chloride  Test.  (Ober- 
mayer's  Keagent).  This  reagent  has  the  advantage  of 
keeping  indefinitely.  It  is  prepared  by  dissolving  2  grams 
of'  solid  ferric  chloride  in  500  cc.  of  concentrated  hydro- 
chloric acid.     (Sp.  gr.  1.19). 

To  equal  parts  of  urine  and  the  above  reagent  add  a 
little  chloroform.  Shake  frequently  but  not  too  violently 
(otherwise  an  emulsion  may  be  formed).    The  chloroform 


V)A  V«^  UAN^_  C^N^  SW^^^a*AlJ^Jv 


will  become  more  or  less  blue  by  the  indigo  formed,  in  pro- 
portion to  the  indican  originally  present.  According  to 
the  depth  of  the  blue  color  it  may  be  designated  as  little, 
much  or  copious. 

Oxalic  Acid.  This  is  usually  found  in  combination  with 
lime  in  the  form  of  calcium  oxalate.  The  crystals  are  of  small 
size  and  appear  in  the  form  of  dumb  bells  and  octahedra.  They 
occur  normally  in  greater  amount  in  herbivorous  than  in  omni- 
vorous urine.  They  greatly  increase  after  eating  such  vegetables 
as  tomatoes,  fresh  beans,  beet-root,  asparagus,  apples  grapes, 
honey,  and  after  the  use  of  rhubarb,  senna,  squills,  etc.  Another 
source  of  oxalic  acid  in  the  body  is  incomplete  oxidation  of  carbo- 
hydrates and  proteids  or  retarded  metabolism.  It  is  therefore  a 
result  of  mal-assimilation  and  is  found  in  dyspepsia,  diabetes  mel- 
litus,  etc.  The  long  continued  excretion  of  an  excess  of  oxalate  of 
lime  frequently  irritates  the  kidneys,  producing  albuminuria  and 
grave  nervolis  disturbances  and  may  lead  to  the  formation  of 
calculi.     (Fig.  18.)    '  •■ 

Acetone.  Normal  lirine  may  contain  traces  of  acetone  but 
it  occurs  in  excessive  quantities  as  a  pathological  condition.**  It 
is  found  in  many  of  the  fevers,  certain  forms  of  cancer,  in 
starvation,  and  in  diabetes,  when  it  indicates  an  advanced  form 
of  the  disease.  It  is  associated  ^ith  an  increased  proteid  meta- 
bolism and  is  looked  upon  as  a  product  of  proteid  decomposition 
with  deficient  oxidation. 

Lieben-Kalfe  Test.  Dissolve  1.3  grams  (20  grains) 
of  potassium  iodide  in  4  cc.  (1  dram  of  liquor  potassae. 
Boil  in  test  tube,  after  which  gently  pour  the  urine  on 
its  surface.  A  yellow  precipitate  between  the  two  solu- 
tions indicates  an  affirmative  tejit.  A  more  satisfactory 
,  test  is  to  add  to  the' urine  a. few  crystals  of.  iodine  and  of 
iodide  of  potassiuin  with  some  caustic  potash.  Heat.  Yel- 
'^     ■  low  precipitate — iodoform,  with  its  cha^'acteristic  odor. 

Legal's  Test  for  Acetone.  Add  to  5  cc.  of.  the  urine 
some  fresh  aqueous  solution  of  sodium  nitrbp'russi'de  fol- 
lowed by  a  little  ammonia,  or  sodium  hydrate  solution'' 
which  give's  a  red  color  disappearing  on  boiling...  Add.suf- 


*  If  pathological  ui'ine  is  not  available,  a  .small  amount  of  acetone 
may  be  added  to  the  urine  for  laboratory  tests.    .  ' 


31 

ficieut  glacial  acetic  acid  and  a  purple  or  violet  red  color 
results.     Compare  with  ereatinin. 

These  tests  are  not  always  satisfactory  when  applied 
to  the  ordinary  urine.  Greater  accuracy  is  claimed  if  the 
urine  is  distilled  and  the  tests  applied  to  the  distillate. 

Sternberg's  Test.  Acidulate  the  suspected  fluid  with 
a  few  drops  of  phosphoric  acid,  and  then  add  small  quan- 
tities of  solution  of  copper  sulphate,  and  of  iodine  in 
potassium  iodide  (Lugol's  Solution)  ;  when  acetone  is 
present  a  brow^n  cloudiness  appears ;  on  heating,  the  liquid 
is  decolorized,  and  a  grayish-white,  pulverulent,  volumin- 
ous precipitate  appears,  which  contains  iodine  and  copper 
in  organic  combination.  The  precipitate  is  almost  insol- 
Tible  in  water.  Alcohol  affords  a  similar  reaction,  but  only 
after  prolonged  boiling,  and  gives  a  sparing  precipitate. 

Frommer's  Test  for  Acetone.  To  10  cc.  of  urine  in 
a  test  tube  add  1  gram  of  solid  potassium  hydroxide ;  be- 
fore the  latter  is  dissolved  add  10  to  12  drops  of  salicylic 
aldehyde  (made  by  dissolving  1  part  of  salicylic  acid  in 
10  parts  of  absolute  alcohol).  Heat  the  mixture  to  about 
70°  C.  In  the  presence  of  acetone,  there  is  formed  a  scar- 
let red  ring.  According  to  Frommer,  even  the  minutest 
amount  of  acetone  will  give  this  reaction  and  no  other 
constituent  of  the  urine  will'  give  this  color — not  even 
diaceticacid.  .  The  reafction  is  explained  as  follows:  One 
molecule  of  salicylic  aldehyde  combines  with  one  molecule 
of  acetone  to  form  oxybenzol-acetone.  This,  in  the  pres- 
ence of  strong  alkalies,  forms  dioxy-dibenzol-acetone.  The 
alkaline  salts  of  this  compound  are  intensely  red. 

Urobilin  is  commonly  regarded  as  the'  most  important 
coloring  matter  in  the  urine.  There  is  some  'evidence  that  it 
represents  a  reduced  form  of  bilirubin,  on^  of  the  pigments  of 
the  bile.*  Urobilin  is  more  readily  obtaimed  from  highly  colored 
urines,  (f fevers,  etc.). 

Test.  The  ordinary  test  wath  an  alcoholic  solution 
of  zinc  may  be  simplified  in  the  following  manner:  10  cc. 
of  urine  are  acidified  with  2  drops  of  HCl  and  shaken  with 

*  A  small  amount  of  an  alcoholic  extract  of  the  feces  added  to  the 
urine  will  usuaiiy  give  favorable  tests  for  urobilin. 


s„JXSi  \>^^    '>^ 


\Nh<>J\ 


,£a^ 


32 

2  ee.  of  chloroform.  After  the  separation  of  the  liquids^ 
2  cc.  of  the  chloroform  layer  are  tested  Math  4  cc.  of  a 
solution  of  1  gram  of  crystallized  acetate  of  zinc  in  a  liter 
of  95%  alcohol  (shelf  reagent).  At  the  junction  of  the 
two  layers  the  green  fluorescent  ring  characteristic  of  uro- 
bilin will  appear,  and  on  shaking,  a  fluorescence  which  is 
rose-colored  by  reflected  light  will  be  distinguished 
throughout  the  liquid. 

Another  test  is  to  add  ammonia  to  the  urine  until 
distinctly  alkaline,  filter,  and  to  the  filtrate  add  a  little 
10%  chloride  of  zinc  solution.  A  green  fluorescence  should 
appear,  and  if  examined  with  the  spectroscope,  a  Charac- 
teristic band  should  occur.     (See  Fig.  16.) 

Leucin  and  Tyrosin  are  pathologic  constituents  of  urine. 
They  are  normal  products  of  pancreatic  digestion  and  under 
ordinary  conditions  are  carried,  after  absorption,  to  the  liver, 
where  they  disappear,  presently  undergoing  decomposition. 
When  present  in  the  urine  these  bodies  ^ire  usually  considered 
pathognomonic  of  acute  yellow  atrophy  of  the  liver,  although 
they  are  likewise  stated  to  be  present  in  the  urine  in  certain 
rare  cases  of  acute  phosphorus  poisoning  associated  with  hepatic 
atrophy,  due  to  typhoid  fever,  etc. 

Phenol.  According  to  Tereg  and  Munk  the  horse  excretes 
in  the  urine  about  10  grams  of  tribromphenol  in  24  hours.  The 
tribromphenol  is  equivalent  to  3  grams  of  phenol  daily.  Great 
importance  is  laid  by  these  observers  on  the  excretion  of  phenol, 
a  process  which  is  suspended  during  intestinal  complaints,  par- 
ticularly colic,  and  is,  according  to  them  and  others,  a  cause  of 
the  rapid  death  in  these  affections,  produced  by  the  toxic  effect 
of  the  unexcreted  phenol.  The  production  of  phenol  in  the 
healthy  body  is  greatly  influenced  by  diet,  being  largest  on  rye 
and  hay,  one  part. peas  and  two  parts  oats,  and  on  hay  alone;  it 
is  smallest  on  rye  alone,  and  next  smallest  on  oats  and  hay.  Sal- 
kowski  is  inclined  to  regard  the  excretion  of  3  grams  of  phenol 
daily  as  too  high. 


:j3 


ABNORMAL  SUBSTANCES  FOUND  IN  THE  URINE. 

Albumin.  The  presence  of  this  substance  in  the  urine  is 
regarded  as  pathologic.  There  is,  however,  in  the  urine  of  some 
individuals  apparently  enjoying  perfect  health,  minute  traces  of 
albumin  sometimes  present,  and,  unless  these  traces  persist,  are 
not  to  be  regarded  as  serious.  If  present  in  any  considerable 
quantity,  it  must  be  regarded  as  distinctly  abnormal.  Albumi- 
nuria is  the  term  applied  when  albumin  occurs  in  notable  quan- 
tity in  the  urine.  The  principal  form  of  albumin  present  is 
serum-albumin,  in  addition  there  may  be  serum-globulin,  acid 
albumin,  albumose,  and  peptone. 

The  amount  of  albumin  in  the  urine  may  be  increased:  1. 
By  food  rich  in  albumin;  2.  Suppression  of  cutaneous  perspira- 
tion, as  by  colds,  burns,  or  cutaneous  diseases ;  3.  Pulmonary  and 
cardiac  diseases  attended  with  dyspnoea,  cyanosis,  valvular  dis- 
eases of  the  heart ;  4.  Febrile  and  inflammatory  diseases,  as  ma- 
larial, eruptive,  typhus  and  typhoid  fevers,  croup,  diphtheria, 
erysipelas,  rheumatism,  gout,  peritonitis,  meningitis,  etc. ;  5.  By 
lesions  or  prostration  of  the  nervous  system,  especially  when  at- 
tended by  diminished  temperature  and  arterial  tension,  as  from 
grief,  fear,  injury,  pressure;  6.  Pressure  as  from  tumors,  preg- 
nancy, etc. ;  7.  Cachexias,  as  from  cancer,  syphilis,  scrofula,  sep- 
ticemia ;  8.  Hydremia  and  ailments  that  disturb  the  vascular  ten- 
sion; 9.  Chorea,  convulsions,  exacerbations  of  febrile  and  other 
diseases;  10.  Diseases  of  genito-urinary  organs,  as  Bright 's  dis- 
ease, cystitis,  hemorrhage,  abscess,  etc. ;  11.  Medicines,  such  as 
copaiba,  cubebs,  turpentine,  some  emetics  and  drastic  cathartics, 
some  anesthetics,  coffee,  many  metallic  salts,  poisoning  by  hydro- 
gen arsenide,  carbon  protoxide,  carbon  dioxide,  phosphorus, 
iodine,  iodoform,  etc. 

The  loss  of  organic  material  (albumin)  disturbs  nutrition, 
the  blood  becomes  more  aqueous ;  it  sometimes  produces  an  anas- 
arca which  is  the  result  of  hydremia  and  of  anuria. 

Albuminuria  may  be  caused:  1.  By  an  alteration  in  the  renal 
transudation ;  2.  By  certain  changes  in  the  blood ;  3.  By  disturb- 
ance of  the  circulation. 

Renal  Changes.     Lesions  of  the  dialysiug  portions  of   the 


34 

convo- 


kidney,  especially  of  the  glonierule ;  the  epithelium  of  the 
luted  tubules  may  also  be  essential  in  the  production  of  albu- 
minuria. These  cells  normally  prevent  the  albumin  from  filtering 
through  with  the  other  elements  of  the  plasma.  Albuminuria  is 
also  dependent  upon  renal  lesions,  sometimes  primary  as  in 
nephritis,  sometimes  consecutive  as  in  alteration  in  the  blood  or 
a  disturbance  in  the  circulation.  Acute  nephritis,  chronic  neph- 
ritis, fatty  or  amyloid  degeneration  interfere  with  the  process  of 
dialysis. 

Bacteria  may  exercise  a  traumatic  action  upon  the  renal 
epithelium  and  cause  desquamation  or  degeneration,  obstruct  the 
vessels,  modify  blood  pressure,  or  by  the  excretion  of  soluble 
products  irritate  the  parts. 

Changes  in  the  Blood.  The  dialysing  membrane  cannot 
stand  with  impunity  any  adulteration  of  the  blood.  The  passage, 
in  the  kidney,  of  any  such  substance  as  biliary  pigment  in  icterus ; 
glucose  in  diabetes ;  poisons,  such  as  alcohol,  lead  or  mercury,  or 
toxic  gases,  render  the  urine  albuminous. 

Subcutaneous  injections  of  solutions  of  extractives  (leucin, 
ty rosin,  creatinin,  xauthin  and  hypoxanthin)  cause  degeneration 
of  the  epithelium  of  the  kidneys  and  albuminuria.  The  subcu- 
taneous injection  of  tincture  of  cantharides  causes,  in  a  few  min- 
utes, the  production  of  an  albuminous  exudate  in  the  glomerules. 
Although  the  limit  of  saturation  of  the  blood  plasma  by  albumin 
may  be  unknown,  it  is  none  the  less  evident  that  a  superabun- 
dance of  the  substance  (albumin)  in  the  vessels  causes  albumi- 


nuria. 


The  existence  of  a  physiologic  albuminuria  is  still  doubtful  in 
the  domestic  animals ;  for  Frohner,  who  has  examined  the  urine 
of  a  number  of  healthy  horses,  has  found  only  two  cases  in  which 
albumin  was  present.  In  man  it  has  boen  demonstrated  to  be  due 
to  severe  muscular  exercise,  slight  cold,  and  nitrogenous  diet. 

Disturbance  of  Circulation.  Too  great  a  variation  in  blood 
pressure  will  cause  albuminuria. 

Renal  emboli,  section  of  vaso-motor  nerves  of  the  kidney, 
medullary  lesions,  etc.,  cause  albuminuria  by  causing  an  active 
congestion  of  the  kidney.  Venous  stasis,  organic  affection  of  the 
heart,  of  the  liver,  presence  of  fetus,  etc.,  cause  albuminous  urine. 

The  urine  may  be  less  fluid.    Bacteria  may  frequently  cause 


35 

thromboses,  emboli,  edemas,  anemias,  etc.,  from  vaso-motor  trou- 
bles changing  simultaneously  the  filter,  the  liquids  which  filter,, 
with  regard  to  pressure  and  velocity. 

Albuminous  urine  is  usually  of  light  color  and  low  specific 
gravity.  It  may  occasionally  be  dark  and  dense,  due  to  other  in- 
gredients, or  to  concentration. 

Under  particular  conditions  of  fatigue  or  disease  albumin 
may  appear  in  the  urine. 

Temporary  albuminuria  is  sometimes  induced  by  a  cold 
bath,  especialty  in  persons  prone  to  kidney  disease,  and  it  has 
been  observed  after  excessive  muscular  exercise,  as  in  the  urine 
of  soldiers  after  a  prolonged  march. 

Any  cause  which  leads  to  an  increased  blood  pressure  in  the 
kidneys  tends  to  induce  albuminuria,  and  many  of  the  cases  in 
which  it  is  the  result  of  disease  may  be  traced  to  this  cause.  Al- 
buminuria is  a  constant  accompaniment  of  the  nephritis  follow- 
ing scarlet  fever  and  may  occur  to  a  less  extent  in  pneumonia, 
typhoid  and  diphtheria.  It  may  also  occur  in  diabetes,  and  is 
then  a  highly  unfavorable  symptom. 

In  every  case  the  urine  must  be  clear  before  testing,  by 
filtering  it  carefully;  also  take  the  specific  gravity.*  In  addi- 
tion to  the  suspected  urines  make  control  tests  on  the  normal  for 
comparison. 

Heat  Test.  Heat  about  5  cc.  of  the  urine  to  the  boil- 
ing point  in  a  test  tube.  Note  the  slightest  turbidity.  If 
present  it  will  be  due  to  albumin  or  earthy  phosphates.  In 
horse  urine  the  precipitate  may  be  due  to  driving  off  CO, 
and  precipitation  of  lime,  etc.,  not  phosphates.  Add 
slowly  a  few  drops  of  acetic  acid  (or  nitric).  If  due  to 
the  phosphates  the  urine  becomes  clear,  if  the  turbidity 
remains  it  is  albumin.  Care  must  be  taken,  in  the  addition 
of  the  acid  after  boiling,  to  note  the  effect  after  each  drop 
is  added  and  to  go  on  adding  until  there  is  no  doubt  that 
the  urine  is  distinctly  acid.  If  only  a  trace  of  albumin  is 
present  and  too  much  acid  is  added  the  albumin  may  be 
converted  into  acid  albumin  and  remain  in  solution.  Heat 
does  not  coagulate  acid  albumin.  Add  a  little  acetic  acid 
to  dissolve  any  phosphates  and  heat  again. 

*  If  a  pathological  urine  is  not  available,  a  little  blood  serum  added 
to  the  normal  urine  will  give  satisfactory  tests. 


8(1 

Another  Heat  Test.  Fill  a  test  tube  about  one-third 
full  of  water  and  boil  it.  Add  a  few  drops  of  the  sus- 
pected urine.  If  albumin  is  present  a  cloudiness  or 
coagulum  will  appear,  according  to  the  amount  of  albumin 
present. 

Heller's  Cold  Nitric  Acid  Test.  Pour  some  of  the 
urine  gently  upon  the  surface  of  some  nitric  acid  in  a  test 
tube.  A  ring  of  white  coagulum  occurs  at  the  junction  of 
the  two  fluids.  If  the  quantity  of  albumin  is  small,  the 
coagulum  may  not  occur  for  a  few  minutes.  A  brown  zone 
will  frequently  be  seen  at  the  point  of  contact  due  to  the 
action  of  the  acid  upon  the  coloring  matters  of  the  urine, 
but  it  does  not  give  any  turbidity  unless  albumin  be 
present. 

Millard-Eobert 's  or  Nitric  Magnesian  Test.  The 
reagent  is  as  follows:  Nitric  Acid,  1  part;  Sat.  Sol.  Mag- 
nesium Sulphate,  5  parts.    Use  as  in  the  preceding  test. 

Picric  Acid  Test.  (Johnson's).  Fill  the  test  tube 
half  full  of  urine.  Slightly  incline  the  tube  and  gently 
pour  down  its  side  about  2  cc.  of  a  saturated  solution  of 
picric  acid,  so  that  it  may  come  in  contact  with  the  upper 
layer  of  the  urine.  Place  the  tube  in  an  upright  position. 
A  layer  of  coagulated  albumin  will  appear  at  the  line. of 
junction.  The  coagulation  of- albumin  takes  place  at  once^ 
and  is  thus  not  easily  mistaken  for  precipitated  urates, 
which  require  soihe  time  for  their  precipitation  and  dis- 
appear on  the  application  of  heat. 

Ferrocyanide  Test.  To/ 2  ec.  of  acetic  acid  in  a  test 
tube  add  4  cc.  of  a  5%  solution  of  potassium  ferrocyanide. 
Mix  them  and  add  10  cc.  of  urine.  A  precipitate  will  ap- 
pear if  albumin  be  present.    No  heat  is  required. 

A  test  equally  as  good  is  that  proposed  by  Zouchloss 
in  which  potassium  sulphocyanide  is  substituted  for  the 
ferrocyanide. 

Sugar  in  the  Urine.  Dextrose  or  glucose  exists  in  the  blood 
from  0.8  to  1.25  parts  per  1000.  When  a  greater  amount  than 
3  parts  per  1000  exists,  the  excess  is  excreted  through  the  kidneys. 
It  is  maintained  by  many  that  a  trace  of  dextrose  is  normally 
present  in  the  urine  and  may  appear  in  slightly  larger  quantities 


37 

transitorily  without  pathologic  significauee.  The  presence  of 
small  quantities  of  sugar  in  the  urine  is  designated  as  glycosuria ; 
in  larger  quantities  it  is  known  as  diabetes  mellitus.  The  former 
condition  if  habitual,  is  unnatural,  and  may  terminate  in  the 
latter. 

These  diseased  conditions  do  not  necessarily  point  to  dis- 
eases of  the  kidney  or  urinary  organs,  but  rather  to  the  liver. 
The  kidney,  in  ridding  itself  of  this  product  (dextrose),  becomes 
irritated,  and  this  irritation  extends  down  the  entire  canal,  and 
we  thus  have  a  real  polyuria  produced. 

Sugar,  in  sufficient  quantity  to  react  to  ordinary  tests,  is 
found  in  the  urine  physiologically  during  pregnancy  and  lac- 
tation ;  in  infants  under  two  months  old ;  in  old  persons  living 
largely  upon  starchy  and  saccharine  food.  Pathologically  in 
diabetes  mellitus;  in  impeded  respiration  from  pulmonary  dis- 
eases; in  impeded  hepatic  circulation  (functional  and  organic 
diseases  of  the  liver)  ;  in  diseases  of  the  central  nervous  system 
(general  paresis,  epilepsy,  dementia,  puncture  of  the  fourth  ven- 
tricle) ;  in  intermittent  and  typhoid  fevers,  by  the  action  of  cer- 
tain poisons,  as  carbon  monoxide,  arsenic,  chloroform  and  curare  : 
in  abnormally  stout  persons. 

The  persistent  excretion  of  easily  recognizable  quantities  of 
sugar  constitutes  diabetes.  The  quantity  of  urine  is  often  enor- 
mously increased,  as  much  as  10000  cc.  being  passed  in  24.  hours, 
by  man.  The  specific  gravity  is  high,  varying  from  1025  to  1050. 
The  color  is  usually  pale,  from  the  dilution — not  diminution — of 
the  urinary  pigments. 

The  presence  of  albumin,  interferes  with  the  tests  for  sugar 
and  must,  in  all  cases,  be  removed  by  the  addition  of  acetic  acid 
and  heat.  The  urine,  after  being  filtered,  may  then  be  used  for 
the  sugar  tests. 

The  particular  property  of  glucose,  which  is  utilized  for 
its  detection,  is  its  action  as  a  reducing  agent — its  disposition  to 
absorb  oxygen.  In  this  property  it  difPers  strikingly  from  sac- 
charose or  common  cane  sugar. 

Principle  of  the  Copper  Tests.  If  a  little  copper  sulphate 
and  an  excess  of  a  solution  of  caustic  potash  be  added  to  a  solu- 
tion of  glucose,  a  clear  blue  solution  results.  Without  the  glucose, 
the   alkali   would  precipitate    the    pale  blue    cupric    hydrate. 


38 

CuOoHs ;  and  if  the  mixture  were  boiled  this  blue  precipitate 
would  be  reduced  to  a  black  precipitate  of  Cupric  Oxide,  CuO,. 
The  clear  blue  solution  containing  glucose,  however,  when  boiled, 
changes  from  transparent  blue  to  opaque  yellow,  and  speedily  de- 
posits a  yellow,  ultimately  red,  precipitate  of  cuprous  oxide  CuO. 
When  the  quantity  of  sugar  is  large  the  change  is  immediate. 
When  small  the  reaction  takes  a  few  minutes  for  its  completion. 

Fehling's  Solution.  Solution  A.  34.64  grams  of  pure  crys- 
talline copper  sulphate  are  powdered  and  dissolved  in  500  cc.  of 
distilled  water.  Solution  B.  Sodio-potassium  tartrate  (Rochelle 
Salts)  173  grams.  Pure  caustic  potash  125  grams.  Add  enough 
distilled  water  to  make  500  cc.  When  using  take  equal  parts  of 
Solutions  A  and  B.  (If  diluted  with  5-10  vols,  of  water  the  test 
is  said  to  be  more  sensitive). 

Place  some  Fehling's  Solution  in  a  test  tube  and  boil 
it.  If  no  yellow  discoloration  takes  place  it  is  in  good 
condition.  Add  a  few  drops  of  the  suspected  urine  and 
boil.  If  the  mixture  suddenly  turns  to  an  opaque  yellow 
or  red  color,  the  presence  of  a  reducing  sugar  is  indicated. 
Normal  horse  urine  probably  because  of  its  pyrocatechin 
usually  changes  the  color  of  the  copper  solution,  but  this 
does  not  indicate  sugar. 

Benedict's  Modification  of  Fehling's  Test.  Greater 
delicacy  and  accuracy  are  claimed  for  this  test.  •  There  are 
two  solutions  as  in  Fehling's.  The  first  is  prepared  by 
dissolving  34.65  grams  of  cupric  sulphate  in  a  small 
amount  of  water  and  made  up  to  500  cc.  The  second  by 
dissolving  100  grams  of  anhydrous  sodium  carbonate  and 
173  grams  of  Rochelle  salt  dissolved  in  water  and  made  up 
to  500  cc.  These  solutions  should  be  preserved  separately 
in  rubber-stoppered  bottles  and  mixed  in  equal  volumes 
when  needed  for  use.  This  is  done  to  prevent  deteriora- 
tion. 

To  2  cc.  of  Benedict's  solution  in  a  test  tube  add  6  cc. 
of  distilled  water  and  not  more  than  7  to  9  drops  of  the 
urine  under  examination.  Boil  the  mixture  vigorously  for 
15  or  30  seconds  and  allow  it  to  cool  to  the  room  tempera- 
ture. If  sugar  is  present  in  the  solution  a  precipitate  will 
form  which  is  often  hluish-green  or  green  at  first,  especially 


v<>N\  ^Jv/V\*JI: 


^ 


39 

if  the  percentage  of  sugar  is  low,  and  which  usually  be- 
comes yellowish  on  standing.  If  the  sugar  present  exceeds 
0.06%  this  precipitate  generally  forms  at  or  below  the 
boiling  point,  whereas,  if  less  than  0.06%  of  sugar  is  pres- 
ent the  precipitate  forms  more  slowly  and  generally  only 
after  the  solution  has  cooled. 

Benedict  has  further  modified  the  test  by  making  a 
single  solution  which  does  not  deteriorate  upon  long 
standing.     The  formula  is  as  follows: 

Cupric  Sulphate  17.3  grams 

Sodium  Citrate  173.0      " 

Sodium  Carbonate   (anhydrous)     100.0      '' 
Distilled  Water  to  1000.0  cc.  ^ 

With  the  aid  of  heat  the  sodium  citrate  and  carbonate 
are  dissolved  in  about  600  cc.  of  water.  Pour  (through  a 
folded  filter  if  necessary)  into  a  glass  graduate  and  make 
up  to  850  cc.  The  cupric  sulphate  is  dissolved  in  about 
100  cc.  of  water  and  made  up  to  150  cc.  The  carbonate- 
citrate  solution  is  poured  into  a  large  beaker  and  the  cupric 
sulphate  solution  is  added  slowly,  with  constant  stirring. 
The  mixed  solution  is  ready  for  use,  and  does  not  deterior- 
ate upon  long  standing. 

The  procedure  is  as  follows :  To  5  cc.  of  the  reagent  in 
a  test  tube  add  not  more  than  8  drops  of  the  urine  to  be 
examined.  The  fluid  is  then  boiled  vigorously  for  one  or 
two  minutes  and  then  allowed  to  cool  spontaneously.  In 
the  presence  of  dextrose  the  entire  body  of  the  solution 
.  will  be-  filled  with  a  precipitate,  which  may  be  red,  yellow 
oi*''^4/ywi!^M'cbiOTf  cl^5tfen*ding  upon  the  amount  of  the  sugar 
present.  If  no  dextrose  is  present,  the^Mution'-will  eitlf^slRi^ 
%.viS^'*ig';^^i68i^ft^^^|^i^iKill  show  a  very  faint  turbidity, 

due"  to  preciprtlrFed  urare^  '    '  ''t^'  <  ■'^ •  - '  %*^*\.. 

Trommer's.Test.  To  4  or  5  cc.  of  urine,  in  a  test  tube, 
add  one-Half  its  volume  of  20%  caustic  potasli.  and  2  or  3 
drops  of  a  solution  of  copper  sulphate  (1-10).  Heat  to 
the  boiling  point.  If  sugar  be  present  a  yellowish  or  red- 
dish precipitate  is  thrown  down,  the  sugar  having  re-  ' 
dueed  the  cupric  hydrate  to  cuprous  oxide. 

Bismuth  Test.     Put  equal  quantities'  of  urine  and 


?  <^s^  \^ 


.\  \'<\ 


0^ 


40 


20%  solution  of  caustic  potash  in  a  test  tube  and  add  a 
pinch  of  subnitrate  of  bismuth.  Boil  the  mixture  and 
if  glucose  be  present  the  powder  turns  black.  Albumin 
and  sulphur  also  reduce  bismuth  and  must  be  removed  if 
the  test  is  to  be  reliable.  Bismuth  is  more  or  less  black- 
ened with  normal  horse  urine. 

Ny lander's  Reagent.  A  solution  is  made  of  bismuth 
subnitrate  2  grams ;  Rochelle  salts  4  grams ;  potassium 
hydroxide  10  grams;  and  distilled  water  to  100  cc.  The 
urine  is  heated  to  boiling  and  a  few  drops  of  this  alkaline 
solution  of  bismuth  added,  and  on  continuing  the  boiling, 
if  sugar  be  present,  the  mixture  turns  black.  The  test  is 
delicate,  as  little  as  0.025%  of  glucose  can  be  detected. 

Phenylhydrazine  Test.  (Modified  from  Kowarsky). 
To  5  drops  of  pure  phenylhydrazine  and  10  drops  of 
glacial  acetic  acid  in  a  test  tube  is  added  1  cc.  of  a  10% 
solution  of  sodium  chloride.  After  shaking  the  mixture, 
3  cc.  of  the  urine  are  added  and  the  test  tube  heated  for 
two  minutes  or  longer.  The  fluid  is  then  allowed  to  cool 
slowly.  If  the  sugar  contents  exceed  0.5%  the  precipitate 
of  glucosazone  takes  place  in  about  two  minutes.  Small 
quantities  of  albumin  do  not  hinder  the  reaction,  but  are 
precipitated  by  boiling.  After  an  hour  or  more  examine 
the  precipitate  microscopically.  A  10  9r  solution  of  sodium 
hydroxide  has  been  recommended  as  a  substitute  for  the 
sodium  chloride  solution  and  greater  delicacy  is  claimed 
for  the  reaction. 

The  glucosazone  crystals  will  have  the  form  typical 
of  skeins  of  wool,  which  will  generally  be  double,  whereas 
other  crystals  precipitated,  such  as  those  of  glycuronic 
acid,  are  irregularly  formed.  As  little  as  0.005 7f  (one- 
fortieth  grain  per  ounce)  of  glucose  can  be  detected  in 
urine  by  the  phenylhydrazine  reaction. 

Uric  acid,  urea,  xanthine,  and  creatinin  in  no  way 
simulate  the  reaction  of  glucose  with  phenylhydrazine. 
The  only  bodies  which  can  offer  confusion  are  glycuronic 
acid  and  its  derivatives. 

FermentationTest.  Robert  'sDifferentialDensity Method. 
Take  the  specific  gravity  of  the  urine  before  adding   the- 


W^^^    VMtaIV      j^lb^iSlv  ^^^iJ-v  l-'^^.^    ^^^ 


41 


yeast  and  record  it.  Mix  well  2  fluid  ounces  (60  cc.)  of 
urine  with  ^  cake  of  compressed  yeast  in  a  bottle.  Set 
aside  for  24  hours  in  a  moderately  warm  place.  After  the 
fermentation  filter  and  take  the  specific  gravity  again  and 
subtract  from  that  taken  before.  Each  degree  of  the  re- 
mainder represents  one  grain  of  glucose  to  the  fluid  ounce. 
Multiply  by  0.219  to  get  the  percentage.  Thus:  Specific 
gravity  before  fermentation,  1035 ;  specific  gravity  after 
fermentation,  1015.  1035 — 1015^20  degrees  of  density 
lost,  or  20  grains  of  sugar  to  the  fluid  ounce.  This  test 
is  conclusive  as  to  the  presence  of  sugar,  though  it  is  not 
absolutely  accurate  as  to  quantity. 


Bile  in  the  Urine.  In  a  number  of  pathologic  conditions  the 
elements  of  the  bile  are  excreted  in  the  urine.  The  bile  pigments, 
bilirubin  and  biliverdin,  may  occur  along  with  the  bile  salts, 
sodium  glycocholate  and  taurocholate,  or  the  bile  salts  alone  may 
be  present. 

Urine  containing  the  bile  pigments  is  colored  a  yellow  brown 
or  brownish  green.  It  forms  an  intense  yellow  froth  on  agita- 
tion. It  stains  paper  or  linen  a  permanent  yellow.*  The  bile 
pigments  are  found  in  jaundice,  functional  disorders  of  the  liver 
(acute  and  chronic  biliousness)  ;  organic  diseases  of  the  liver 
apart  from  jaundice  (carcinoma,  amyloid  disease,  cirrhosis)  ; 
diseases  of  the  spleen;  fever;  hemolytic  diseases  (anemia,  leuco- 
cythemia  and  scurvy). 

Gmelin's  Test.  (Nitric  acid  containing  nitrous 
acid).t  Place  a  few  drops  of  the  suspected  urine  in  a 
white  porcelain  dish  and  near  them  a  few  drops  of  the 
impure  nitric  acid;  let  the  fluids  run  together  and  the 
usual  play  of  colors  is  observed. 

Put  some  urine  in  a  test  tube  and  carefully  pour  in 
some  of  the  yellow  impure  nitric  acid,  until  it  forms  a 


*  A  little  omnivorous  or  carnivorous  bile  may  be  added  to  the 
normal  urine  to  demonstrate  the  tests  if  no  icteric  urine  is  available. 

t  Nitrous  acid  may  be  prepared  by  heating  a  little  nitric  acid  to 
which  a  small  amount  of  starch  has  been  added. 


\^ ;^K^  wWV  ^(UW  oil  ,ft<A 


42 


stratum  at  the  bottom.  If  the  bile  pigments  are  present 
at  the  line  of  junction  of  the  fluids,  a  play  of  colors  takes 
place — from  above  downwards — green,  blue,  violet  or  dirty 
red,  and  yellow.  Nearly  all  urines  give  a  play  of  colors, 
but  green  is  the  necessary  and  characteristic  color  to  prove 
the  presence  of  the  bile  pigments. 

Pettenkofer's  Test  for  Bile  Acids.  Take  about  2  cc. 
of  suspected  urine  in  a  test  tube  and  add  4  drops  of  a  10% 
solution  of  the  cane  sugar.  Add  strong  sulphuric  acid, 
drop  by  drop,  cooling  the  tube  in  a  dish  of  cold  water  im- 
mediately after  adding  the  acid.  Not  more  than  2  cc.  of 
the  acid  should  be  used.  Too  much  heat  causes  carboniza- 
tion of  the  sugar  and  the  test  is  ruined.  If  bile  acids  are 
present,  the  fluid  at  first  becomes  opaque,  then  clear  and 
successively  brown,  red  and  purple.  It  may  require  an 
hour  or  more  to  accomplish  the  test.  This  reaction  de- 
pends upon  the  production  of  furfurol  by  the  destruction 
of  the  sugar  when  the  sulphuric  acid  is  used.  Furfurol  in 
turn  combines  with  cholalic  acid,  formed  by  the  action  of 
the  sulphuric  acid  on  the  bile  acids,  giving  the  color. 

Pettenkofer's  test  may  also  be  quite  satisfactorily 
performed  more  quickly  by  putting  a  little  of  the  sus- 
pected urine  in  a  porcelain  capsule,  adding  a  few  drops  of 
a  solution  of  cane  sugar  and  then  a  few  drops  of  strong 
sulphuric  acid,  keeping  the  mixture  cool  to  prevent  car- 
bonization. 

..  tldranszky  has  modified  Pettenkofer's  test  by  using 
furfurol  directly  instead  of  waiting  for  its  formation  by 
the  action  of  the  sulphuric  acid  upon  the  sug^r.  His  pro- 
cedure is  as  follows :  One  cubic  centimeter  of  the  urine  is 
treated  with  one  drop  of  a  D.1%  solution  of  furfurol.  and 
slowly  superposed  upon  1  cc.  of  sulphuric  acid,  care  being 
taken  to  prevent  too  great  heating  of  the  mixture.  A 
purple  color  appears  at  the  plane  of  contact,  that  gradu- 
ally extends  upward  into  the  superposed  solution ;  on 
standing,  the  color  turns  bluish.  In  alcoholic  solution  a 
green  flourescence  is  seen.  The  pigment  gives  a  typical 
spectrum. 

Hay 's  Test.    Use  two  test  tubes ;  in  one  place  a  little 


AiiK.  \A  WJU.  \<A^  (Am    W^oC\^< 


43 


normal  urine  and  in  the  other  some  of  the  suspected  urine_ 
Leave  the  test  tubes  in  the  rack  in  order  to  prevent  agita- 
tion. Drop  a  little  finely  powdered  sulphur  upon  the  sur- 
face of  the  urines.  If  hile  or  biliary  acids  are  present  in 
the  suspected  urine  the  sulphur  will  sink  at  once  to  the 
bottom  of  the  tube,  while  in  the  normal  urine  it  will  re- 
main upon  the  surface  or  but  a  slight  amoimt  may  sink,  if 
the  tubes  are'  not  agitated. 

Chloroform  as  a  test  for  bile  is  quite  satisfactory. 
Agitate  a  few  drops  of  chloroform  with  the  suspected 
urine  in  a  test  tube.  If  bile  be  present  the  chloroform 
becomes  turbid  and  acquires  ayeUowish  hue,  the  depth  of 
which  is  in  proportion  to  the  amount  of  bile  present. 

To  some  of  the  suspected  urine  add  a  little  bromine 
water  {1%).  If  bile  is  present  a  green  ring  should  appear. 
Blood.  Blood  is  sometimes  a  constituent  of  the  urine  m 
disease  It  may  occur  in  two  forms :  1.  As  hematuria,  when  the 
blood  coloring  matter  is  present  in  the  urine  in  combination  with 
blood  corpuscles.  2.  As  hemoglobinuria,  when  no  blood  corpus- 
cles are  present  and  the  blood  pigment  or  hemoglobin  is  m  solu- 
tion in  the  urine. 

Hematuria  mav  have  its  source  (1)  in  the  kidneys  due  to 
injuries;  acute  nephritis;  acute  acerbation  of  chronic  nephritis; 
diseases  of  renal  vessels  (embolism,  thrombosis,  aneurism,, 
stasis)  ;  amyloid  kidney  (very  rarely)  ;  infective  fevers  (small- 
pox, scarlatina,  typhoid  fever,  etc.);  certain  blood  diseases 
(scurvy,  purpura,  hemophilia):  parasitic  diseases  (echmo- 
coccus)  2  The  renal  pelvis  and  ureters  due  to  renal  calculi; 
tuberculosis:  rupture  of  neighboring  abscesses;  parasites^  3. 
The  bladder  due  to  calculi;  cancer  and  other  tumors;  diph- 
theritic cystitis ;  varicose  veins :  injuries.  4.  The  urethra  due 
to  injury  (catheterization,  impaction  of  calculi,  etc.).  5.  Ex- 
traneous discharges  as  the  menstrual  flow,  etc. 

Hemoglobinuria  has  been  observed  in  severe  infectious  dis- 
eases   (tvphoid  fever,  scarlatina,  etc.)  ;  in  conditions  of  blood 
dissolution  (scurw,  purpura,  etc.)  ;  in  skin  burns,  sunstroke,  etc. 
Heller's  Blood  Test  is  made  by  adding  a  little  caustic 
soda  solution  to  some  urine  in  a  test  tube  and    heating. 


i^v^  m-^  W— ^  ^-^  -"^ 


44 

The  precipitated  phosphates  are  colored  reddish  brown 
and  fall  in  a  thick  cloud  to  the  bottom  of  the  tube. 

Almen's  Test.  Add  a  few  drops  of  freshly  made 
5%  alcoholic  tincture  of  guaiacum  to  the  suspected  urine. 
Shake  well  and  add  a  few  drops  of  hydrogen  dioxide  or  of 
old  oil  of  turpentine,  and  the  mixture  will  turn  greenish 
blue.  The  hemoglobin  will  change  the  color  of  the  pre- 
cipitate to  blue.  The  test  will  not  respond  to  a  small  quan- 
tity of  blood. 

Hemin  Test.  If  some  blood  or  sediment  supposed  to 
contain  blood  be  heated  carefully  with  some  glacial  acetic 
acid  and  a  trace  of  sodium  chloride,  and  then  slowly  evap- 
orated in  the  air,  brownish  yellow  rhombic  crystals  of 
hemin  are  found. 

The  spectroscope  and  microscope  are  also  used  in 
blood  tests. 

Melanine.  In  certain  cases  of  melanosis  this  pigment 
appears  in  the  urine,  which,  when  emitted  is  clear,  but  gradually 
becomes  of  a  deep  brown,  or  even  black  color. 

Ordinarily  melanine  exists  in  solution  in  the  urine,  but 
sometimes  in  the  form  of  brownish  or  black  sediment,  recog- 
nizable by  microscopic  examination.  Melanine  may  possess  a 
diagnostic  significance  when  the  melanosis  is  beyond  the  reach 
of  examination  by  eye  or  touch. 

It  may  disappear  from  the  urine  when  the  disease  is  arrested, 
or  it  may  remain  stationary.  Oxidizing  agents,  such  as  chromic 
acid  and  fuming  nitric  acid,  transform  the  principle  melanine, 
causing  gradually  with  the  first  and  immediately  with  the  second 
a  black  coloration.  According  to  Zeller,  the  most  delicate  test  for 
melanine  is  bromine  water.  With  melanine  it  gives  at  first  a  yel- 
low precipitate,  which  gradually  blackens.  Urobilin  gives  a  yel- 
low precipitate  with  the  same  reagent,  but  it  does  not  blacken. 

The  practical  significance  of  this  condition  (melanuria)  is 
greatly  limited  by  the  fact  that  the  urine  may  contain  a  large 
quantity  of  melanine  in  wasting  diseases,  whilst  that  derived 
from  individuals  suffering  from  melanotic  cancer  or  sarcoma  may 
be  entirely  free  from  it.  Senator  has  recently  confirmed  this  view 
by  a  series  of  clinical  observations.  Nevertheless,  as  an  adjunct 
in  diagnosis,  the  tests  given  are  of  undoubted  utility. 


45 


VII. 


QUANTITATIVE  ANALYSIS. 

Centrifugal  Method.  The  centrifuge  as  ordinarily  used  gives 
the  hulk  percentage  of  the  constituents,  this  percentage  being 
arrived  at  arbitrarily  as  a  result  of  numerous  determinations 
upon  normal  urine.  Knowing  the  normal  bulk  percentage  it  is 
not  difficult  to  determine  an  abnormal  or 
pathologic  amount  of  a  substance  as  indi- 
cated by  its  excess  or  deficiency,  and  the 
method  is  therefore  a  convenient  and 
expeditious  one  for  clinical  purposes. 
To  a  beginner,  however,  the  method  as 
commonly  employed,  is  misleading  in 
that  the  bulk  percentage  and  the  true  or 
actual  percentages  are  widely  different. 
Take  for  example  the  phosphates  of  the 
urine.  The  normal  hulk  percentage  as 
given  by  the  centrifuge  is  8%  for  man; 
1%  or  less  for  the  horse.  Whereas,  the 
real  amount  of  phosphates  present  as 
P^Og  in  the  urine  of  man  for  the  whole 
24  hours  (1500  cc.  of  urine)  is  only 
about  3.5  grams,  or  in  1000  cc.  about  2.5 
grams,  or  a  true  percentage  of  0.25%. 
The  true  percentage  may  be  calculated 
from  the  centrifuge  by  giving  to  each  0.1  cc.  of  precipitate  a  de- 
termined value  with  reference  to  the  amount  of  PoO.^  present. 

Each  centrifuge  tube  is  graduated  to  15  cc.  The  first  10  cc. 
are  divided  so  that  each  cubic  centimeter  is  divided  into  10  parts, 
each  part  representing  0.1  cc.  The  remaining  5  cc.  are  for  hold- 
ing the  test  reagents  which  are  added  to  the  10  cc.  of  urine.  The 
first  cubic  centimeter  because  of  the  tapering  end,  will  admit  of 
finer  graduation  than  the  remainder  of  the  tube.  The  first  half 
of  the  cubic  centimeter  is  divided  into  ^/^o  cc.  (or  .025  cc.)  in 
order  that  small  amounts  of  precipitate  may  be  accurately  read ; 
the  second  half  of  the  cubic  centimeter  is  divided  into  V20  cc.  (or 
.05  cc.)  ;  while  the  remaining  9  cc.  are  each  divided  into  Vio  cc. 
(or  0.1  cc.)  as  above  stated.  It  is  convenient  to  take  the  ^/lo  cc. 
as  the  unit  in  the  calculation  of  the  true  percentage.    By  taking 


Fig.  9. 
Hand  Centrifuge. 


46 


Fig.  10.     Aluminum  Tubes 


Sedimentation 
Tube. 


the  average  of  a  great  number  of  control  tests  with  the  burette  it 
has  been  found  that  Vio  cc  of  the  precipitate,  in  the  case  of  the 
phosphates,  represents  0.13  gram  of  P.Og  in  1000  cc.  of  urine. 
If  albumin  is  present,  remove  it  by  adding  a  little  acetic 
acid  and  applying  heat.  Filter  and  test  the  filtrate  for  the  inor- 
ganic constituents. 

With  the  horse  urine  care  must  be  exercised  in  adding  the 
reagents.  The  large  amount  of  carbonates  present  cause  con- 
siderable effervescence,  from  the  liberation  of  the  CO,,  and  some 
of  the  urine  is  likely  to  flow  over  the  tube.  Add  a  little  of  the 
reagent  and  when  the  effervescence  has  ceased  add  a  little  more 
until  the  required  amount  has  been  used.  j 

Phosphates.  Fill  the  graduated  tube  to  the  10  cc, 
mark  with  the  urine  to  be  tested.  Add  1  cc.  of  glacial 
acetic  acid  and  4  cc.  of  5%  uranium  nitrate  solution  to 
reach  the  15  cc.  mark.  Invert  the  tube  several  times  and 
revolve  in  the  centrifuge  for  three  minutes.  If,  after  re- 
volving three  minutes  at  1000  revolutions  per  minute,  the 
precipitate  comes  up  to  the  eighth  0.1  cc.  line  of  the  tube 
(0.8  cc.)  multiply  0.13  by  8  for  the  product,  which  is  1.04 
grams  of  PoOg  in  1000  cc.  If  the  total  amount  of  urine  for 
the  24  hours  is  1400  cc,  the  amount  of  PoOg  present  in  this 
quantity  can  easily  be  calculated  by  the  following  propor- 
tion :  1.04  gm.  P.O.,  :  1000  cc.  : :  X  :  1400.  In  which  the 
value  of  X  is  found  to  be  1.456  grams  of  PoO.,  in  24  hours. 
Make  the  same  determination  with  the  urine  of  the  horse. 


>-    1.1 


47 


Chlorides.  Fill  the  graduated  tube  to  the  10  ec. 
mark  with  the  uriue.  Add  15  drops  or  1  cc.  of  nitric  acid 
to  prevent  precipitation  of  the  phosphates.  Then  fill  to 
the  15  cc.  mark  with  the  silver  nitrate  solution  (1  to  8). 
Invert  the  tube  several  times  to  thoroughly  mix  the  re- 
agents. Revolve  in  the  centrifuge  for  intervals  of  three 
minutes  until  the  precipitate  no  longer  settles.  The  aver- 
age amount  of  precipitate  for  man  is  from  0.5  cc.  to  0.8  cc. ; 
for  the  horse  about  0.5  cc.  The  value  of  each  0.1  cc.  is  1.3 
of  a  gram  of  NaCl  per  1000. 

Sulphates.  These  are  estimated  as  insoluble  salts 
of  barium.  Fill  the  graduated  tube  to  10  cc.  mark  with 
fresh  urine  and  add  5  cc.  barium  chloride  solution,  (4 
parts  barium  chloride,  1  part  hydrochloric  acid,  16  parts 
distilled  water). 

The  amount  of  precipitate  for  man  and  horse  is  about 
.075  cc.  normally.  The  value  of  each  0.1  precipitate  is 
2.4  grams  of  SOg. 

In  the  above  tests  write  down    in    your    notes    the 
amount  '.of  each  constituent  for  .the  24  hours  .considering 
■  '.'.1250  ccr'as  thi?  tptatiamomit  of  iirine  passed. 
Quantitative  Estimation  of  Uric  Acid.    This  estimation  is 
difficult  and  time-consuming  and  is  generally  regarded  as  inex- 
pedient for  clinical  purposes.     The  following,  methods  are  de- 
scribed because  they  are  more  suitable  for  clinical  work  although 
less  accurate  than  the  more  elaborate  methods  of  Salkowski,  Lud- 
wig,  Hopkins  and  others. 

Ruhemann's  Uricometer  for  the  rapid  estimation  of  uric 
acid  consists  of  a  graduated  tube  with  a  glass  stopper.  It  is  a 
colorimetric  method,  in  which  iodine  with  carbon  bisulphide  serve 
as  indicators. 

The  following  directions  accompany  each  apparatus : 
Fill  the  glass  tube  to  the  lowest  mark  S  with  carbon  bisul- 
phide.    (It  is  not  necessary  for  the  tube  to  be  quite  dry,  but 
there  must  be  no  drop  of  liquid  at  the  bottom) . 

The  lowest  part  of  the  convexity .  (double  meniscus)  has  to 
be  even  with  mark  S,  see  diagram. 

Add  a  solution  consisting  of  1.5  gm.  iodine;  1.5  gm.  potas- 
sium iodide;  15  gm.  absolute  alcohol  and  185  gm.  distilled 
water. 


\fr^\       VlW^'^    'A\1^ 

1^  '"  X^'' 


Ju 


48 


-0,211 
-0,215 
-0218 
-0.221 
1225 
1228 
).231 


Fig.  11. 
Huhemann's 
Uricometer. 


Fill  up  so  that  the  base  of  the  upper  arch 
of  the  double  meniscus  is  on  a  level  with  mark 
J  as  shown  in  the  illustration. 

Then  add  the  urine  to  be  examined,  which 
has  to  be  at  a  temperature  of  18°  Centigrade, 
to  the  mark  2.45  (2.6  ecm). 

Close  the  tube  with  glass  stopper  and 
shake  well  when  the  carbon  bisulphide  will  be- 
come a  dark  copper  brown  color. 

After  adding  more  urine  under  continued 
strong  shaking,  the  carbon  bisulphide  will  ab- 
sorb all  free  iodine  and  the  mixture  will  look 
like  urine. 

Slowly  adding  more  urine  will  change  the 
yellow  foam,  created  by  the  shaking,  into 
white  foam. 

The  color  of  the  carbon  bisulphide  will 
turn  pink  after  a  while. 

Should  this  color  remain  the  same  after 
the  apparatus  has  been  shaken  repeatedly  and 
turned  upside  down,  add  another  drop  of 
urine  and  keep  up  the  same  procedure  until 
only  a  slightly  reddish  coloration  of  the  car- 
bon bisulphide  remains. 

Now  shake  again  vigorously  and  the  car- 
bon bisulphide  will  turn  porcelain  white  and 
the  urine  will  look  like  cloudy  whey. 

To  recapitulate: — The  adding  of  urine 
has  to  be  stopped  as  soon  as  the  carbon  bi- 
sulphide shows  only  a  slightly  reddish  tint, 
because  this  will  disappear  entirely  after  re- 
peated shakings.  The  test  is  finished  when  the 
indicator  appears  snow-white,  a  $ign  that  all 
iodine  has  been  neutralized  hy  the  urine. 

To  get  rid  of  the  remaining  foam  move 
the  tube  a  few  times  slowly  to  a  horizontal 
position,  then  open  the  stopper  a  little,  to  al- 
low all  liquid  to  settle  in  the  tube. 

The  proportion  of  uric  acid  is  then  read 
off  the  upper 'scale  (per  thousand  of  urine). 

The  percentage    is  obtained    by  putting 


49 

a  0  in  front.  If  the  upper  meniscus  line  of  the  urine  is  between 
any  of  the  0.1  ccm.  marks,  the  upper  number  should  be  read. 

The  figures  to  the  left  of  the  apparatus  refer  to  the  number 
of  cc.  of  urine  added  to  the  mixture. 

Should  the  urine  contain  less  uric  acid  than  the  apparatus 
will  in  this  way  indicate,  add  the  iodine  solution  to  the  mark  half 
way  between  ^  and  J  and  read  after  each  reaction  the  half  values. 

The  vessel  in  which  the  urine  is  to  be  kept,  must  not  be 
cleaned  with  soda. 

If  the  urine  shows  an  acid  reaction,  it  can  be  used  at  once, 
but  if  it  should  be  alkaline,  it  has  to  be  made  acid  by  adding 
diluted  acetic  acid.    Cloudiness  is  of  no  importance. 

If  the  urine  contains  a  considerable  sediment  of  sodium  urate 
it  should  be  well  shaken. 

Strong  colorations  of  the  urine  do  not  affect  the  action  of 
carbon  bisulphide. 

Traces  of  sugar  and  albumin  do  not  disturb  the  result. 

If  there  is  a  very  large  percentage  of  albumin  or  traces  of 
blood  or  pus,  these  pathologic  substances  have  to  be  coagulated 
by  boiling  and  the  urine  filtered. 

The  apparatus  is  not  satisfactory  for  determining  the  uric 
acid  in  the  urine  of  the  horse. 

(The  uricometer  may  be  purchased  of  Eimer  &  Amend,  New 

York). 

Cook's  Method  of  Estimating  Uric  Acid  hy  the  Centrifuge. 
Place  in  the  graduated  tube  10  cc.  of  urine;  add  to  this  1  gm.  of 
sodium  carbonate  and  1  cc.  of  ammonium  hydrate.  Sliake  until 
the  sodium  carbonate  is  dissolved;  this  precipitates  the  earthy 
phosphates.  Separate  this  precipitate  with  the  centrifugal  ma- 
chine and  decant  the  supernatant  urine  into  another  graduated 
tube.  It  will  be  found  that  the  earthy  phosphates  are  readily 
separated  and  adhere  to  the  bottom  of  the  tube;  this  allows  the 
clear  urine  to  be  readily  poured  off  into  another  tube.  To  the 
clear  urine  now  free  from  phosphates  add  2  cc.  of  ammonium 
hydrate  and  2  cc.  of  ammonio-silver  solution  (made  by  dissolving 
5  gm.  of  silver  nitrate  in  100  cc.  of  water  and  adding  ammonia 
until  the  solution  becomes  clear).  The  addition  of  the  silver 
solution  causes  the  uric  acid  to  be  precipitated  as  the  urate  of 
silver,  a  translucent,  shiny  substance.  Separate  this  precipitate 
with  the  centrifugal  machine  and  pour  off  the  supernatant  urine. 
Add  to  the  ppt.  an  excess  of  ammonium  hydrate,  at  least  15  cc. 
and  mix  thoroughly.  By  this  last  addition  any  of  the  chlorides 
that  may  have  been  precipitated  are  redissolved  leaving  only  a 
pale  urate  of  silver.  Lastly  precipitate  this  urate  of  silver  until 
the  lowest  reading  is  to  be  had.  Each  0.1  cc.  as  marked  on  the 
graduated  tube  indicates  0.001175  gram  of  uric  acid  in  10  cc.  of 


■H' 


50 


urine.  The  amount  of  uric  acid  for  the  24  hours  can  be  calcu- 
lated by  the  following  formula,  supposing  that  1250  cc.  was  the- 
amount  of  the  urine  passed. 


0.001175X5 


10 


•X  1250=0.7343  gm. 


Estimation  of  Uric  Acid  by  Weight.  20  cc.  of  hydrochloric 
acid  are  added  to  200  cc.  of  urine  and  the  mixture  set  aside  for  24 
hours.  Uric  acid  crystals  form  and  collect  on  the  sides  of  the 
vessel.  This  may  be  collected  on  a  weighed  filter  and  washed 
thoroughly  with  water.  The  filter  and  uric  acid  are  dried  and 
weighed  until  there  is  no  further  loss  of  weight.  The  weight  of 
the  filter  paper  is  deducted  and  the  result  gives  the  amount  of 
uric  acid  in  200  cc.  of  urine,  from  which  the  amount  in  24  hours 
can  be  calculated. 

Urea.  Doremus  Ureometer  Test.  Fill  the  long  arm  an:l 
bend  of  the  ureometer  with  the  hypobromite  solution.  (Hypo- 
bromite  of  sodium  is  prepared  by  mixing  2  cc.  of  bromine  with 


Fig.  13.  Hind's  modification  of 
Doremus  Ureometer. 


23  cc.  of  a  solution  of  caustic  soda,  40  grams  to  100  cc.  of  distilled 
water.    To  this  mixture  add  an  equal  volume  of  distilled  water) . 


51 


With  a  thoroughly  washed  pipette  draw  up  exactly  1  cc.  of  urine 
and  pass  the  pipette  through  the  bulb  of  the  ureoineter  as  far  as  it 
will  go  in  the  bend.  Compress  the  bulb  of  the  pipette  gently  and 
steadily.  The  urine  will  rise  through  the  hypobromite,  and  the 
urea  instantly  decompose,  giving  off  nitrogen  gas.  AVithdraw 
the  pipette  after  the  urine  has  been  expelled,  taking  care  not  to 
press  the  bulb  hard  enough  to  drive  the  air  out  with  the  urine, 
and  read  the  volume  of  gas,  after  allowing  the  froth  to  subside. 

Each  division  mark  on  the  ureometer  indicates  0.001  gram 
of  urea  in  1  cc.  of  urine.  The  quantity  of  urea  voided  in  24  hours 
is  ascertained  by  multiplying  the  result  of  the  test  by  the  number 
of  cc.  of  urine  passed  during  that  period. 

The  COo  resulting  from  the  decomposition  of  the  urea  is  ab- 
sorbed by  the  excess  of  soda  in  the  hypobromite  solution,  and 
nitrogen  is  evolved.  37.1  cc.  of  moist  nitrogen  gas  measured 
under  the  ordinary  conditions  of  experiment  may  be  taken  to  rep- 
resent 0.1  gram  of  urea.  1  gram  of  urea  corresponds  to  13.72 
grams  of  muscular  tissue. 

Albumin.  Esbach's  Method.  Fill  the 
tube  with  the  suspected  urine  to  the  letter  U, 
then  add  the  reagent  to  the  letter  R.  (The 
reagent  is  made  by  mixing  10  grams  of  picric 
dcid  and  20  grams  of  citric  acid  and  adding 
enough  w^ater  to  make  1  liter.  It  is  said  that 
acetic  acid  may  be  substituted  for  the  citric 
with  as  good  results).  Invert  the  tube  a 
number  of  times  so  that  the  contents  may  be 
thoroughly  mixed.  Close  the  tube  tightly 
with  the  rubber  stopper  and  set  aside  for  24 
hours,  after  which  the  amount  of  dried  albu- 
min in  one  liter  of  urine  can  be  read  in  grams 
on  the  tube.  The  percentage  is  obtained  by 
dividing  by  ten.  Thus,  if  the  coagulum 
stands  at  3,  the  urine  contains  three  parts  of 
albumin  per  thousand,  or  0.3%.  If  the  albu- 
min is  very  abundant  (above  four)  the  urine 

should  be  diluted  to  obtain  an  accurate  result. 

0.5  per  thousand  of  albumin  cannot  be 

mated  by  this  method. 


u^^ 


Fig.  14. 
Esbach's 
Albumin- 
ometer. 

Less  than 
accurately  esti- 


^^k!    .)b\\0  -^  '^'^^  /^^^  -  ^^■/'\ 


liM^-Xv 


jiKv., -o^ini-- ■"i)^^'^ "'    ^' "'"^r* 


14^    ,bMo-.o^ih^**^^ 


t^       .^'t  i»  "'■''/* 


52 

Caution.  The  coagulable  substance-  found  in  the 
urine  of  Bright 's  disease  is  a  mixture  of  ' '  serum-albumin ' ' 
and  ' '  serum-globulin  "  or  "  para-globulin. ' '  Saturation  of 
the  urine  with  crystallized  magnesium  sulphate  precipi- 
tates the  serum-globulin,  leaving  the  serum-albumin  in 
solution. 

Centrifuge  Test.  Fill  the  graduated  tube  with  urine 
up  to  the  10  cc.  mark;  add  5  cc.  of  Esbach's  reagent.  In- 
vert the  tube  several  times  in  order  to  thoroughly  mix  the 
fluids.  Place  the  tube  in  the  centrifuge  and  revolve  for 
three  minutes.  Read  off  the  amount  of  the  precipitate; 
each  0.1  cc.  is  the  equivalent  of  0.21  of  a  gram  of  dry  al- 
bumin per  1000  cc.  of  urine. 

Ferrocyanide  Test  with  Centrifuge.  The  percentage 
tube  is  filled  to  the  10  cc.  mark  with  the  urine  to  be  tested. 
Add  1  cc.  of  acetic  acid  and  4  cc.  of  a  5%  solution  of 
potassium  ferrocyanide.  Proceed  as  in  the  preceding  test. 
Each  0.1  cc.  of  precipitate  is  the  equivalent  of  0.21  gram 
of  dry  albumin  per  1000  cc.  of  urine. 

Sugar.  Einhorn's  Fermentation  Saccharometer. 
Determine  the  specific  gravity.  Add  10  cc.  of  the  sus- 
pected urine  to  90  cc.  of  distilled  water.  Put  in  a  flask, 
add  about  one  gram  of  compressed  yeast  and  agitate 
choroughly.  Pour  10  cc.  of  the  mixture  into  the 
bulb  of  the  saccharometer  and,  by  inclining  the 
apparatus,  the  fluid  will  displace  the  air  in  the 
cylinder  and  remain  there  by  atmospheric  pres- 
sure. Be  sure  that  no  air  bubbles  remain  in  the 
cylinder.  It  is  always  well  to  test  a  normal  urine 
at  the  same  time  and  in  the  same  way  as  a  con- 
trol. The  mixture  of  normal  urine  with  yeast 
will,  on  the  following  day,  have  only  a  small 
bubble  on  the  top  of  the  cylinder.  If,  in  the  sus- 
pected urine,  there  is  also  present  at  the  top  of 
the  cylinder  a  small  bubble,  no  sugar  is  present; 
but  if  there  is  a  much  larger  volume  of  gas  (CO2) 
it  is  certain  that  the  urine  contains  sugar.  The  appar- 
atus should  remain  in  a  moderately  warm  place.  As  a 
result  of  fermentation  the  sugar  is  broken  up  into  alcohol 


53 


and  CO2.  The  changed  level  of  the  fluid  in  the  cylinder 
shows  that  the  reaction  has  taken  place  and  indicated  by 
the^  numbers  the  approximate  quantity  of  sugar  present. 
The  -scale  on  the  tube  is  empirical  and  indicates  directly 
the  percentage  in  the  urine. 

Shieb's  Test  for  Sugar  in  the  Urine. 
Solution  No.  1. 

Ammonium  Sulphate   (purest)  1.2  grams.          r--- 

Copper  Sulphate  (purest)  -         2.6  grams. 

<        Distilled  Wate]^  «   -  -         -       50.    cc. 

Solution  No.  2. 

Caustic  Potash  C.  P.  -         -         20  grams. 

Distilled  Abater  -         -  -      -         50  cc. 

Dissolve  and  when  cool,  add 

Glycerine  •  -         -         -         -  .      50  cc. 

Ammonia  water  0.9G0  sp.  gr.  300  cc. 

Add  No.  1  to  No.  2  and  dilute  the  w^ole  to  500  cc. 
with  distilled  water.  Stopper  securely  and  shake  till 
thoroughly  mixed. 

Heat  one  dram  of  this  solution  in  a  test  tube  to  boil- 
ing. Add  the  urine  drop  by  drop,  at  slow  intervals,  boil- 
ing after  each  addition  until  the  blue  color  has  been  dis- 
charged and  the  fluid  has  a  light  amber  color  or  is  colorless. 

If  the  solution  is  decolorized  by  3  minims  of  urine  it 
contains  9  to  10  grains  of  sugar  per  oz. 

If  the  solution  is  decolorized  by  4  minims  of  urine  it 
contains  7  to  8  grains  of  sugar  per  oz. 

If  the  solution  is  decolorized  by  5  minims  of  urine  it 
contains  5  to  6  grains  of  sugar  per  oz. 

If  the  solution  is  decolorized  by  6  minims  of  urine  it 
contains  4  grains  of  sugar  per  oz. 

If  the  solution  is  decolorized  by  7  minims  of  urine  it 
contains  3  grains  of  sugar  per  oz. 

If  the  solution  is  decolorized  by  9  minims  of  urine  it 
contains  2  grains  of  sugar  per  oz. 

If  the  solution  is  decolorized  by  10  to  17  minims  of 
urine  it  contains  1  grain  of  sugar  per  oz. 

If  the  urine  contains  more  than  10  grains  of  sugar  to 


54 


the  ounce  it  must  be  diluted  with  an  equal  quantity  of 
water,  and  the  number  of  grains  per  ounce  multiplied  by 
two. 

A  very  similar  preparation,  accurately  compounded, 
is  on  the  market  under  the  name  of  Whitney's  Reagent. 
It  is  for  sale  by  the  Norwood  Chemical  Co.,  105  W.  40th 
St.,  New  York. 

The  following  table  gives  the  amounts  of  sugar    in 

analytical  testing  with  Whitney 's  Reag^t :    X  » 

If  reduced  by        It  contains  to  the  oz.  /   Percentage. 


1  minim 

16  grains 

or  more 

3.33 

2  minims 

8  grains 

1.67 

3       " 

5.33 

grains 

1.11 

4      " 

4 

0.83 

5       " 

3.20 

0.67 

6      '' 

2.67 

0.56 

7  .    " 

2.29, 

0.48 

8       " 

2 

0.42 

9       " 

1.78 

0.37 

10       " 

1.60 

(1 

0.33 
0.21) 

Quantitative  Test  hy  Benedict's  Method.  In  the 
quantitative  method  Benedict  uses  three  separate  solutions. 
The  Cupric  sulphate  solution  and  the  Alkaline  tartrate 
solution  are  the  same  as  those  already  described  for  his 
qualitative  test,  (see  p.  38).  The  third  solution  is  the 
Potassium  ferro-thiocyanate  solution  and  is  made  up  as 
follows : 

Potassium  ferrocyanide         -         -         15.    grams 
Potassium  thiocyanate  -         -         62.5      " 

Anhydrous   sodium   carbonate     -         50.        " 
Water  up  to  .         .         -         .       500.    cc. 

The  three  solutions  should  be  preserved  separately  in 
rubber-stoppered  bottles  to  prevent  deterioration  and 
mixed  in  equal  volumes  when  needed  for  use. 

Application.  To  30  cc.  of  Benedict's  solution  in  a 
small  beaker  add  from  2.5  grams  to  5  grams  of  anhydrous, 
sodium  carbonate  and  heat  the  mixture  to  boiling  over  a 


55 


wire  gauze  until  the  carbonate  has  been  brought  into  solu- 
tion. (Where  the  urine  is  diluted  or  there  is  a  low  per- 
'centage  of  sugar  use  the  5  grams  of  the  sodium  carbonate). 

Place  the  urine  under  examination  in  a  burette  and 
run  it  into  the  hot  Benedict  Solution  rather  rapidly  until 
the  formation  of  a  heavy  chalk-white  precipitate  is  noted 
and  the  blue  color  of  the  solution  lessens  perceptibly  in  its 
intensity.  From  this  point  in  the  determination  from  2 
to  10  drops  of  the  urine  should  be  run  rather  slowly  into 
the  boiling  Benedict  Solution  at  one  time,  boiling  the 
solution  vigorously  for  about  15  seconds  after  each  addi- 
tion. Complete  reduction  of  the  copper  is  indicated  here 
as  in  Fehling's  original  method,  by  the  complete  disap- 
pearance of  all  Hue  color.  The  end-point  here,  however, 
is  very  sharply  defined,  contrary  to  the  conditions  in  the 
older  method. 

To  prevent  the  annoying  bumping  which  often  inter- 
feres with  the  titration,  a  medium-sized  piece  of  washed 
absorbent  cotton  may  be  introduced  into  the  solution. 
This  cotton  may  be  stirred  about  through  the  solution  as 
the  titration  proceeds  and  the  bumping  thus  eliminated. 

Calculation.  Thirty  cubic  centimeters  of  Benedict's 
Solution  is  completely  reduced  by  0.073  gram  of  dextrose. 
If  Y  represents  the  number  of  cubic  centimeters  of  urine 
necessary  to  reduce  the  30  cc.  of  the  solution  M:e  have  the 
following  proportion  Y  :  0.073  ::  100  :  .Y  (percentage  of 
dextrose). 

Ehrlich's  Diazo-reaction.  This  test  has  been  recom- 
mended for  the  diagnosis  of  typhoid  fever  in  man.  It  is 
stated  that  the  reaction  will  also  occur  in  some  other  dis- 
eases, but  in  spite  of  this  fact  the  test  is  of  value  as  an  aid 
in  the  diagnosis.     Two  solutions  are  prepared  as  follows : 

1.  Sulphanilic  acid  2  gms. 
Hydrochloric  acid               50  cc. 
Distilled  water                 1000  cc. 

2.  A.  0.5 ^{   solution  of  sodium  nitrite. 

In  performing  the  test,  50  parts  of  No.  1  and  1  part 
of  No.  2  are  mixed,  and  equal  parts  of  this  mixture    and 


'56 


of  the  urine  in  a  test  tube  are  rendered  strongly  alka- 
line with  ammonia.  If  the  reaction  be  positive,  the 
solution  assumes  a  carmine-red  color,  which  on  shaking- 
must  also  appear  in  the  foam.  Upon  standing  24  hours 
a  greenish  precipitate  is  formed.  The  test  must  not  be 
considered  positive  unless  a  distinct  coloration  extends 
to  and  includes  the  foam  on  shaking. 


VIII. 

VOLUMETRIC  METHODS. 

Chlorides.  Mohr's  method.  Principle:  If  silver  nitrate  be 
added  to  a  solution  containing  sodium  chloride,  neutral  potas- 
sium chromate,  and  an  alkaline  phosphate,  the  chloride  is  first 
precipitated,  then  the  chromaje,  and  lastly,  the  phosphate.  The 
formation  of  the  red  silver  chromate  indicates  the  complete  pre- 
cipitation of  the  chloride. 
Sblutioijs  required : 

.  1.     Standard  soluti'on  o:^  silver  nitrate : 
Fused  silver  nitrate  29.075  grams. 
Distilled  water  to  make  1000  cc. 
2.,    Saturated  solution  neutral  potassium  chromate: 
•  Neutral  potassium  chromate  10  grams. 

Distilled  water  to  make  100  cc. 
Process.  The  urine  should  not  be  high  colored, 
and  should  be  free  from  aibumin  or  excess  of  uric  acid 
or  mucus.  Dilute"  10  cc.  of  the  urine  with  100  cc.  of  dis- 
tilled water.  Fill  a  burette  with  the  silver  solution  to 
the  zero  mark.  Drop*it  slowly  into  the  urine,  stir  it  well 
and  occasionally  carry  a  drop  of  the  mixture  so  as  to 
come  in  contact  with  a  little  of  the  chromate  solution  in 
an  evaporating  dish.  Test  in  this  way  until  the  first  trace 
of  .orange  color  appears  in  the  chromate  solution.  Make 
sure  that  the  precipitation  of  the  chlorides  is  complete  by 
adding  another  drop  of  the  silver  solution  from  the 
burette.* Read  off  the  am£)unt  of  silver  solution  used  and 
calculate  the  result. 


cr  -C 


n     ^    CW<A<^«jX 


57 

Example : 

Quantity  of  urine  in  24  hours,       1250     cc. 

Silver  solution  used,  -  7.5  cc. 

1  cc.  silver  solution  equals  .01  gram  NaCl 

.01  X  7.5 

X  1250  =  9.375  grams. 

10 

Make  two  or  three  determinations  and  take  the  aver- 
age for  your  final  result.  Do  the  same  in  the  phosphate 
and  sulphate  tests. 

If  the  urine  of  the  horse  is  very  dark  colored,  it  may 
be  filtered  through  animal  charcoal  to  make  it  light 
colored.  Some  of  the  chlorides  may  be  held  by  the  char- 
coal and  thus  diminish  the  amount  in  the  urine  tested; 
or  the  urine  may  be  diluted  with  an  equal  volume  of  dis- 
tilled water  and  the  result  multiplied  by  two. 

Gravimetric  MetJiod. 

A  more  accurate  method  is  the  following:  If  the  urine 
is  high  colored,  and  contains  albumin,  or  excess  of  uric  acid  and 
mucus,  they  must  be  removed.  To  do  this,  measure  10  cc.  of  the 
urine  into  a  platinum  capsule,  add  2  grams  of  pure  potassium 
nitrate,  evaporate  to  dryness,  and  ignite  at  a  dull  red  heat  to 
destroy  organic  matter.  When  cool,  treat  the  residue  with  hot 
water  and  filter.  Acidulate  the  filtrate  with  dilute  nitric  acid, 
neutralize  with  carbonate  of  lime  and  proceed  as  above. 

Estimation  of  Phosphoric  Acid.  (Estimated  as  PoOg).  By 
uranium  acetate  or  nitrate.  This  method  is  based  upon  the  fact 
that  when  a  solution  of  acetate  or  nitrate  of  uranium  is  added  to 
a  solution  containing  soluble  phosphates,  sodic  acetate  and  free 
acetic  acid,  all  the  phosphoric  acid  will  be  precipitated  as  phos- 
phate of  uranium.  This  precipitate  is  of  a  light  yellow  color, 
insoluble  in  acetic,  but  soluble  in  hydrochloric  acid.  The  point 
of  completion  of  the  phosphoric  reaction  may  be  ascertained  by 
placing  with  a  glass  rod  a  drop  of  the  yellow  mixture  in  contact 
with  a  drop  of  potassium  ferrocyanide  solution  upon  a  white 
plate  or  filter  paper.  As  soon  as  there  is  the  slightest  excess  of 
the  uranium  solution  after  the  phosphates  have  been  satisfied,  a 
brown  precipitate  will  result  in  the  mixture  due  to  formation  of 
ferrocyanide  of  uranium.  The  cochineal  solution,  noted  below, 
serves  as  a  better  indicator  for  the  horse  urine  than  the  ferro- 
cyanide.   The  following  solutions  are  used : 


58 

1.  A  Solution  of  Cochineal  prepared  by  boiling  40  grams 

of  cochineal    in  800  cc.  of    water.     When    cool   add 
200  cc.  of  alcohol  and  filter. 

Or  a  solution  of  potassium  ferrocyanide. 

Potassium  ferrocyanide,  -  5  grams. 

Distilled  water  -  100  cc. 

2.  Solution  of  Sodium  Acetate. 

Sodium  acetate,  -  100  grams. 

Acetic  acid,        -  -  100  cc. 

Distilled  water  to  make  1000  cc. 

The  following  procedure  is  of  use  in  standardizing  the  uran- 
ium solution.' 

3.  Standard  Solution  of  Disodic  Hydric  Phosphate,  made  by  dis- 
solving 10.0845  grams  of  the  crystallized  salt  in  water  and  diluting  to  1 
liter.  Each  cc.  of  this  solution  contains  .002  of  a  gram  of  P^Oj.  In  5  cc. 
there  is  0.1  gram  of  P2O5. 

4.  Uranium  Acetate.  Since  this  cannot  be  obtained  sufficiently 
pure  to  be  weighed  out  and  used  directly  we  make  a  solution  of  it  of 
indefinite  strength  and  standardize  it  with  the  other  solutions. 

It  has  been  found  best  to  make  the  solution  of  uranic  acetate  of 
such  a  strength  that  each  cc.  will  precipitate  .005  of  a  gram  of  P.Oj. 
Consequently  every  2  cc.  of  the  uranium  acetate  should  be  made  equal 
to  every  5  cc.  of  the  sodic  phosphate,  or  upon  adding  20  cc.  of  the 
uranium  acetate  to  50  cc.  of  the  sodium  phosphate  and  then  touching 
the  plate  or  paper  which  has  been  moistened  with  the  cochineal  solution 
with  the  drop  of  the  mixture,  we  should  just  get  the  green  color. 

Put  50  cc.  of  the  sodium  phosphate  with  5  cc.  of  the  sodium  acetate 
solution  into  a  beaker.  To  this  add  slowly  from  the  burette,  the 
uranium  acetate,  testing  occasionally,  for  the  color  on  the  paper.  Sup- 
pose that  on  the  addition  of  8  cc.  from  the  burette,  the  color  is  obtained, 
then  8  cc.  of  the  uranium  acetate  are  as  strong  as  20  should  be,  and  for 
every  8  cc.  of  the  uranium  solution  that  we  have,  12  cc.  of  water  should 
be  added.  If  it  should  require  more  than  20  cc.  to  produce  the  color,  the 
uranium  solution  must  be  concentrated  by  evaporation  or  more  of  the 
solid  salt  added.    The  solution  has  now  been  graduated  or  standardized. 

Application.  50  cc.  of  the  clear  urine,  with  5  cc.  of 
the  sodium  acetate  solution  are  poured  into  a  beaker  and 
heated;  to  this  the  uranium  acetate  is  slowly  added  from 
the  burette.  The  mixture  is  constantly  st,irred  with  a  glass 
rod,  which  should  be  applied  frequently  to  the  cochineal 
solution.  As  soon  as  the  green  color  is  obtained  the 
process  is  completed.  Eead  off  from  the  burette  the 
amount  of  uranium  acetate  solution  used. 

Example.  Urine  in  24  hours  equals  1180  cc.  Solu- 
tion uranium  acetate  used  equals  24.3  ee. 


lo-O^l  '^   a^&-^w^ 


59 

.005  X  24.3  cc. 

X  1180  =  2.86  grams  P2O5. 

50 

A  shorter  and  approximately  accurate  method  based 
on  the  above  is  as  follows :  To  10  cc.  of  the  urine  in  a 
test  tube  add  1  cc.  of  sodium    acetate    and  1  cc.  of  the 
cochineal  solution  and  boil.     Add  5%  uranium    nitrate 
solution,  1  minim  at  a  time,  boiling  after  each  addition, 
until  the    mixture    turns    green.     Each    minim    of    the 
uranium  solution  represents  0.046  gram  of  PoOg  in  1000  cc. 
of  urine.     Multiply  .046  by  the    number    of    minims  re- 
quired to  produce  the  green  color,  and  the  product  will 
represent  the  amount  in  grams  of  PgOg  per  liter  in  the 
given  sample  of  urine. 
Estimation  of  Total  Sulphuric  Acid.   (Estimated  at  SO3). 
Dissolve  30.5  grams  of  pure  crystallized  chloride  of  barium    in. 
some  distilled  M-ater  and  dilute  to  1  liter.    Each  cc.  of  this  solu- 
tion will  equal  .01  gram  of  SO3.     A  dilute  solution  of  sodium 
or  magnesium  sulphate  will  also  be  required. 

Application.  50  cc.  of  clear  urine  are  poured  into  a 
beaker,  acidified  quite  strongly  with  hydrochloric  acid, 
and  heated  over  the  flame.  Let  the  solution  boil  for  about 
ten  minutes,  the  lamp  is  then  removed  and  the  barium 
chloride  is  allowed  to  flow  slowly  from  the  burette  into 
the  beaker  and  it  must  continue  to  flow  as  long  as  the 
precipitate  is  seen  to  increase.  The  precipitate  is  allowed 
to  subside,  then  more  of  the  barium  chloride  is  added,  and 
this  process  repeated,  until  no  further  precipitate  is  pro- 
duced. Much  time  and  labor  will  be  saved  by  filtering  a 
feAV  drops  of  the  solution  now  and  then,  and  allowing 
these  to  drop  into  the  test  tube  containing  some  of  the 
dilute  sodium  or  magnesium  sulphate.  As  soon  as  an  ex- 
cess of  the  barium  chloride  has  been  added,  a  precipitate 
will  appear  in  the  test  tube.  Read  off  from  the  burette  , 
the  amount  of  barium  chloride  used ;  each  cc.  of  which  will 
indicate  .01  of  a  gram  of  SO3  in  each  cc.  of  urine,  and 
from  this  the  total  amount  may  be  calculated. 

Example :  Urine  2000  cc.    Amount  of  barium  chloride 
solution  25  cc. 


I,  U  MA 


&i\^^'^^_^-  n-3^  jHc-K     f^ 


.?^f> 


60 
.01  X  25 


X  2000  =  10  grams  SO.. 

50 

Relation  of  Urinary  Constituents  in  Normal  Human  Urine. 

There  is  normally  a  direct  proportion  within  narrow  limits,  be- 
tween the  solid  substances  of  the  urine,  which  it  is  desirable  to 
keep  in  mind. 

Kelation  of  Urea  to  total  solids  50%  or  one-half. 

"  inorganic  matter  to  other  solids  30%. 

"  uric  acid  to  urea  2.5%  or  V^q. 

"  nitrogen  in  urea  to  total  nitrogen  91%. 

"  phosphoric  acid  to  urea  12.5%  or  Vg. 

"  chloride  of  sodium  to  urea  407o. 

"  the  sulphates  to  total  nitrogen  18%. 


IX. 
CHEMICAL  EXAMINATION  OF  URINARY  DEPOSITS. 

There  is  generallj"  a  more  or  less  voluminous  deposit  in  the 
urine  after  standing  for  24  hours;  sometimes  this  deposit  is 
formed  in  the  bladder  and  sometimes  it  forms  after  the  emis- 
sion of  the  urine ;  it  is  important  to  note  this  fact.  Deposits  or 
sediments  should  not  be  confounded  with  calculi.  The  former 
have  a  pulverulent  form  while  the  latter  have  the  appearance  of 
grains  or  granules  of  greater  or  less  size. 

It  is  often  very  easy  to  determine  the  nature  of  the  deposit 
by  a  microscopic  examination,  but  chemical  reagents  in  some 
cases  give  more  precise  results. 

With  the  urine  containing  sediment,  shake  thoroughly  to 
distribute  it,  then  pour  into  a  centrifugal  tube  and  revolve  until 
the  sediment  is  completely  precipitated.  Note  the  reaction  of 
the  urine,  whether  acid  or  alkaline.  Pour  off  the  clear  superna- 
tant fluid.    If  the  urine  is  acid  pursue  the  following  scheme. 

Acid  Urate  of  Soda.  If  the  deposit  is  more  or  less  red,  treat 
with  a  little  boiling  water.  If  the  sediment  is  dissolved  it  is  com- 
posed of  the  acid  urate  of  soda.    Caustic  potash  does  not  dissolve 


61 


it,  hydrochloric  acid  gives  a  crystalline  precipitate,  the  deposit 
will  give  the  murexide  test.    (Page  26). 

Uric  Acid.  If  a  crystalline  deposit,  almost  insoluble  in 
boiling  water,  but  soluble  in  caustic  potash  and  giving  the 
murexide  test,  it  is  uric  acid. 

Cystine,  (very  rare).  The  deposit  is  crystalline,  insoluble 
in  caustic  potash  but  soluble  in  ammonia  and  hydrochloric  acid. 

Pus.  A  white,  dense,  mucus  deposit,  viscid  and  not  often 
mixing  with  the  urine;  caustic  potash  renders  it  more  viscid; 


Plate  II. 
Various  forms  of  Calcium  Carbonate  Crystals.     (Horse  Urine). 


62 


the  microscope  shows  the  pus  corpuscles, 
in  acid  urines. 


This  deposit  is  rare 


Blood.  Red  deposit ;  the  tincture  of  guaiac  with  hydrogen 
dioxide  gives  a  blue  color.  The  spectroscope  gives  the  charac- 
teristic lines  in  the  spectrum.  The  hemin  test  will  show  the 
characteristic  crystals.  When  blood  is  present  the  urine  is  gen- 
erally albuminous. 

Some  deposits  not  very  abundant  and  having  no  special 
chemical  reaction  may  be  recognized  under  the  microscope. 

Neutral  or  Alkaline  Urine. 

Alkaline  Urates.  More  or  less  reddish  deposits  easily  soluble 
in  boiling  water  and  giving  the  murexide  test. 

Triple  Phosphate  or  Calcium  Phosphate.  A  white  deposit, 
insoluble  in  boiling  water,  soluble  in  acetic  acid,  does  not  give 
the  murexide  test;  acidified  with  nitric  acid,  the  molybdate  of 
ammonia  solution  gives  a  yellowish  precipitate  in  the  cold. 

Calcium  Oxalate.  White  deposit  insoluble  in  boiling  water, 
also  in  caustic  potash  and  acetic  acid.  Soluble  in  hydrochloric 
acid  or  in  nitric  acid;  does  not  give  the  murexide  test. 

Calcium  Carbonate.  White  deposit  insoluble  in  boiling 
water  or  in  caustic  potash.  Soluble  with  effervescence  in  acetic, 
nitric  or  hydrochloric  acids,  does  not  give  murexide  reaction. 


A 

II 

1                  •'    •             : 

B 

1 

1 

ill 

\\ 

I  ^d!     i  ORANGE  YELLOVJJ  gIREEN  BLUE 

VtOLETj 

c. 

i 
1 

i 

i 

Fig.  16. 
A. — Spectrum  of  Oxy-hemoglobin.    B. — Spectrum  of  Reduced  Hemo- 
globin.   C. — Spectrum  of  Urobilin. 


63 


Pus,  (more  frequent  in  alkaline  and  albuminous  urine).  A 
white,  dense,  mucus-like  deposit,  not  mixing  readily  with  the  rest 
of  -^the  urine.  Caustic  potash  renders  it  more  viscid,  the  micro- 
scope shows  pus  globules. 

Blood.  Determined  by  the  guaiacum  test,  spectroscope,  mi- 
croscope, or  hemin  test. 

From  the  pathologic  point  of  view  the  presence  of  urinary 
sediments  generally  indicate  an  alteration  of  the  secretions. 


X. 


Microscopical  Examination  of  Urine.  If  an  immediate  ex- 
amination is  desired  the  centrifuge  may  be  used  to    cause    the 

sediment  to  separate  from 
the  urine ;  otherwise  the 
urine  is  set  aside  for  some 
hours  in  a  cool  place  when 
the  sediment  gradually 
settles  to  the  bottom.  The 
supernatant  liquid  is  pour- 
ed off  and  by  means  of  a 
pipette.  camel's  hair 
brush  or  a  wire  loop  a 
small  portion  of  the  cle- 
FIG.  17.    Uric  Acid  Crystals.  ^^^-^    -^  transferred    to     a 

slide  and  examined. 

The  deposits  may  be  divided  into  unorganized  and  organized 
sediments. 

Unorganized  Sediments.  In  unorganized  sediments  two  con- 
ditions may  be  considered :  1st,  the  urine  is  acid.  2nd,  the  urine 
is  alkaline. 

In  acid  urine  look  first  for  crystals  of  uric  acid  diverse  in 
form  and  reddish  brown  in  color.  (Fig.  17).  Second,  crystals  of 
acid  urate  of  soda,  yellowish  or  red.  The  crystals  are  badly 
formed.  Third,  crystals  of  oxalate  of  lime  also  found  in  alka- 
line urine.  (Fig.  18.)  Fourth,  crystals  of  hippuric  acid.  Fifth, 
crystals  of  calcium  sulphate.  (4  and  5  are  met  with  rarely  in 


64 


acid  urine).  Sixth,  calcium  phos- 
phate. Make  sure  of  the  identifi- 
cation by  reference  to  charts. 

In  alkaline  urine  look  first  for 
ammonio-magnesium  phosphates, 
(Fig.  19),  (triple  phosphates).. 
Second,  bicalcium  phosphate  crys- 
tals. Third,  tricalcium  or  amorph- 
ous phosphate  crystals.  Fourth, 
crystals  of  sulphate  of  lime.  Fifth, 

crystals  of  oxalate  of  lime,   (also 
FIG.  18.    Calcium  Oxalate.  f^^^^^^     -^^     ^^-^     ^^^-^^^^         g-^^j^^ 

crystals  of  urate  of  ammonium  in  the  form    of   yellow  colored 
spheres. 


Fig.  19.     Triple  Phosphates. 

Crystals  of  cystin,  leucin  and  tyrosin  are  sometimes  en- 
countered in  acid  or  alkaline  urines. 

Organized  Sediments.  These  are  brought  more  plainly  into 
view  if  the  preparations  are  stained,  although  this  is  not  uni- 
versally practised.  Organized  sediments  may  be  divided  into  (a) 
histologic  elements,  (b)  microbic  elements. 

Of  the  histologic  elements  there  are  frequently  encountered, 
1st,  epithelial  cells  from  the  bladder,  from  the  vagina  and  from 
the  ureter,  their  presence  has  no  special  significance.  If,  how- 
ever, they  are  present  in  considerable  quantity,  a  lesion  of  these 
parts  may  be  suspected.  2nd,  cells  from  the  pelvis  of  the  kid- 
ney, generally  an  indication   of   renal    affection.     3rd,    cancer 


65 


Fig.  20. 
Micrococcus  Ureae. 


cells,  which  have  a  special  significance, 
ith,  hyaline  or  granular  casts,  seen  more 
easily  in  stained  preparations;  they  are 
encountered  frequently  in  albuminous 
urine;  their  presence  indicates  a  mild 
form  of  nephritis.  5th,  epithelial  hem- 
orrhagic or  wax  casts,  of  which  the 
hemorrhagic  are  the  most  frequent,  they 
are  markedly  colored  and  easily  recog- 
nized on  account  of  the  hemoglobin  they 
contain ;  they  contain  fine  granulations 
which  are  not  blood  corpuscles  but  possible  fragments  of  them, 
they  indicate  a  severe  form  of  nephritis.  6th,  cylindroid  ele- 
ments drawn  out  in  an  irregular  and  somewhat  ribbon-like 
form ;  they  are  the  product  of  the  secretion  of  the  urinary  epi- 
thelium. In  the  human  race  they  may  be  found  in  cases  of 
scarlatina  and  generally  in  certain  forms  of  nephritis.  In  ex- 
amining for  casts  it  is  desirable  to  examine  the  urine  immedi- 
ately after  emission,  as  they  generally  disintegrate  rapidly  and 
disappear.  7th, 
blood      corpuscles 

more  or  less  ere-  /&^'^^i^^(^%,\  /.^-^ 
nated  but  easily 
recognizable  b  y 
their  yellowish 
tint;  the  presence 
^f  blood  corpuscles 
in  the  urine  indi- 
cates      a       hemor-  ^^''-  ^l-    ^^d  blood  corpuscles.   A.    Normal. 

rhage  either  in  the  bladder  or  in  the  kidney.  8th,  pus  corpuscles 
which  may  have  crenated  borders,  granular  contents  and  appear 
quite  refractive,  a  drop  of  dilute  acetic  acid  will  render  the  nuclei 
visible.  Pathologically  the  pus  corpuscles  indicate  a  suppuration 
of  some  portion  of  the  urinary  tract.  In  the  majority  of  cases  it 
is  impossible  to  say  if  the  pus  comes  from  the  bladder  or  kidney. 
In  cases  of  cystitis  of  the  neck  of  the  bladder,  the  last  portion  of 
the  urine  passed  contains  no  pus,  while  if  this  trouble  be  in  the 
kidney  this  last  portion  will  contain  pus.  This  fact  is  of  some 
Tise  in  diagnosing  cystitis  of  the  neck  of  the  bladder.     9th,  sper- 


GQ 


matozoa  may  be  pres- 
ent but  are  easily  rec- 
ognized by  their 
elongated  form.  10th, 
mucus  may  be  fre- 
quently present,  but 
has  no  very  great 
pathologic  signifi- 
cance. 

Glycogen  Cells.  If 
the  urinary  sediment 
is  stained  with  dilute 
L  u  g  0  1  '  s  Solution, 
there  will  be  seen,  in 
many  specimens  of 
human  urine,  a  num- 
FiG.  22.  Red  blood  corpuscles  crenated.  ber  of  epithelial  cells 
present  with  more  or  less  of  their  interior  stained  a  deeper 
brown  like  glycogen,  than  in  other  cells  that  may  be  present. 
As  yet,  nothing  has  ben  determined  in  regard  to  their  import- 
ance. 

Amyloid  or  Amylaceous  Bodies..  These  are  small  circular 
or  oval  bodies  which  give  the  starch  reaction  with  an  iodine  solu- 
tion. Just  what  their  significance  is,  is  not  known.  Virchow  first 
described  them.  They  have  been  found  in  various  tissues  and 
excretions,  including  the  urine  normal  as  well  as  pathological. 
In  diseases  of  the  kidneys,  Veitz  and  Wederhake  found  that  these 
amyloids  afford  us  important  indications ;  but  that  in  affections 
of  the  bladder  they  are  present  in  increased  quantity.  Weder- 
hake therefore  suggests  that  the  amyloid  bodies  have  a  certain 
value  in  differential  diagnosis.  The  absence  of  the  bodies  in 
pathological  urine  is  against  catarrhal  disease  of  the  bladder; 
whilst  their  presence,  especially  if  numerous,  in  urine  which  ap- 
pears to  contain  renal  elements  only,  indicates  that  the  bladder 
is  also  affected.     (Dixon  Mann). 

The  urinary  sediment  stained  with  a  dilute  Lugol's  solution 
show  these  bodies,  when  present,  varying  in  color  from  a  blue  to 
a  deep  blue  black  color. 

Urinary  Casts.    Although  probably  observed  earlier,  Henle 


rr 


*#■!*! 


p^^#i 


B 


^MM 


E 


^ 


K 


S^^ 
'Z?®^' 


.^^&4 


9  ^^, 


"^^  £1 


P:^. 


.#  J^^ 


"^^p^Jf^T^H 


I^J^ 


Plate  III. 

Urinary  Sediment.  A.  Uric  Acid.  B.  Ammonium  Urate.  C.  Acid 
Sodium  Urate.  D.  Urea  Nitrate.  E.  (1)  Leucin  and  (2)  Tyrosin.  F. 
Cystin.  G.  Ammonio-Magnesium  or  triple  phospliate.  H.  Calcium  Phos- 
phate. I.  Calcium  Oxalate.  J.  Blood  Corpuscles.  K.  Pus  and  Mucus.  L. 
Hemin  Crystals.  M.  (1)  Hyaline  Casts,  (2)  Granular  Casts.  N.  Epi- 
thelial Casts  and  Cells.  O.  (1)  Waxy  Casts;  (2)  Casts  with  Blood  Cor- 
puscles;  (3)  Casts  with  Fat  Globules.     (After  Simon). 


68 


is  credited  in  1842,  with  first  carefully  describing  the  casts 
moulded  in  the  tubules  of  the  kidneys.  Among  earlier  views, 
casts  were  considered  as  being  composed  of  coagulated  fibrin; 
as  products  of  the  secretion  of  the  epithelium  of  the  tubules;  as 
transformed  or  disintegrated  epithelial  cells  and  as  products 
from  the  blood.  A  view  quite  commonly  accepted  is  that  casts 
are  the  products  of  the  coagulation  of  albuminous  material.  The 
fact  that  the  presence  of  casts  in  the  urine  is  usually  accompanied 
by  the  presence  of  albumin  lends 
force  to  this  view;  for  the  more 
abundant  the  albumin,  the  more 
likelihood  is  there  of  finding  casts. 
From  this  standpoint,  then,  casts, 
may  be  regarded  as  albuminous 
exudates  from  the  blood,  with  the 
addition  of  transformed  or  de- 
stroyed epithelium.  In  nearly  all 
cases  where  casts  are  present, 
some  albumin  is  found.  Occasion- 
ally they  are  sometimes  described 
as  being  present  without  the  ap- 
pearance of  albumin.  This  condi- 
tion is  looked  upon  with  some 
doubt ;  for  it  is  commonly  be- 
lieved that  albumin  exists  but  in 
an  amount  too  slight  to  be  detected  by  the  ordinary  chemical 
tests. 

The  presence  of  casts  in  the  urine  is  of  much  diagnostic  im- 
portance ;  if  found  in  any  quantity,  they  indicate  nephritis  par- 
ticularly if  albumin  is  also  present  in  any  amount.  It  is  claimed 
by  some  that  merely  hyperemia  of  the  kidney  will  cause  the 
appearance  of  casts  in  the  urine,  and  that  they  may  sometimes  be 
found  when  the  kidneys  are  perfectly  intact.  Mitchell  states 
that  two  or  three  hyaline  or  one  or  two  small  granular  casts  may 
be  found  in  one  of  every  three  specimens  of  the  twenty-four 
hour's  urine  examined  (human).  Casts  have  been  described  in 
eases  of  gastro-intestinal  catarrh,  in  jaundice,  in  acute  and 
chronic  anemia,  as  well  as  in  nervous  affections  of  different  kinds, 
without  accompanying  inflammation  of  the  kidneys.     There  are 


Fig.  23.  a.  Showing  forma- 
tion of  hyaline  cast  in  tubule, 
b.  Hyaline  cast  with  granular 
deposit,  c.  Granular  cast. 


69 


many,  however,  who  hold  that  easts  are  always  the  products  of 
an  inflammatory  process,  and  are,  therefore,  indicative  of  renal 
inflammation.  Other  evidences  or  symptoms  should  also  be 
taken  into  account. 

Occasional- 
ly it  is  diffi- 
cult to  find 
casts  even 
when  they 
are  known  to 
exist.  Alka- 
line urine  has 
a  tendency  to 
dissolve  them 
At  times 
they  will  not 
settle  for 
hours.  Usu- 
ally if  al- 
lowed to 
stand  s  i  x 
hours,  the 
casts  will  set- 
-le  if  they  are 
present.    The 

centrifuge  will  bring  them  down  in  a  few  minutes.  A  low  power 
of  the  microscope  (150-200  diameters)  may  be  used  in  the  search 
for  casts  and  will  enable  the  observer  to  pass  over  the  field  quite 
rapidly.  To  identify  the  cast  and  its  structure,  a  power  of  400- 
500  diameters  is  required.  More  than  one  specimen  must  always 
be  examined  before  giving  a  positive  statement  as  to  the  presence 
or  absence  of  casts.  False  or  pseudo-casts  have  been  described; 
these  are  believed  to  be  accidental  formations,  while  true  casts  in 
general  indicate  nephritis. 

True  casts  may  appear  in  three  different  sizes  according  to 
the  portion  of  the  tubule  in  which  they  are  formed.  The  smallest 
size  originates  in  the  narrow  tubule.  The  next  in  size  comes  from 
the  convoluted  tubule  of  the  second  order  (no  casts  from  the 
convoluted  tubule  of  the  first  order,  i.  e.,  that   portion    of   the 


Fig.  24.  a.  Blood  cast  and  hyaline  cast  carrying 
blood  cells,  b.  Pus  cast.  c.  Hyaline  cast  carrying  epi- 
thelial cells,  d.  Epithelial  casts.     (Greene). 


70 


tubule  nearest  the  glomerules,  appear  in  tlie  urine,  because  they 
are  too  large  to  pass  through  the  narrow  tubules) .  The  third  and 
largest  are  those  developing  in  the  straight  collecting  tubules. 
It  is  believed  a  prognostic  value  may  be  attached  to  the  size  of 
the  casts  as  well  as  to  their  number.  Casts  from  the  narrow 
tubules  indicate  a  mild  attack;  from  the  convoluted  tubules  a 
severe  form,  especially  in  the  cortex  of  the  kidney;  from  the 
collecting  tubules,  with  the  other  forms  also  present,  a  serious 
condition  of  general  renal  inflammation  or  unfavorable  prognosis. 
The  identity  of  casts  may  generally  be  determined  by  their  uni- 
form width.  They  are  usually  longer  than  they  are  broad,  and 
have  one  well-rounded  extremity  and  well  defined  borders. 

True  casts 
are  of  six  va- 
rieties ;  hya- 
line, epitheli- 
al, blood, 
granular,  fat- 
ty and  waxy 
easts.  In  a 
general  way 
the  first  three 
varieties  are 
found  in  an 
acute  form  of 
n  ep  h  r  i  t  is. 
During  the 
first  few 
weeks  of  the 
inflammation 
the  last  three 
varieties  are 
not  encoun- 
tered. If  the 
acute  condition  passes  into  a  subacute,  then  the  granular  variety 
appears,  at  first  in  small  number,  then  in  larger,  with,  usually, 
a  considerable  number  of  the  hyaline  and  epithelial  casts.  Fatty 
and  waxy  casts  are  always  secondary  products,  and  as  a  rule 
not  found  until  a  nephritis  has  existed  for  some  time. 

Hyaline  casts  are  colorless,  pale,  more  or  less  transparent 


Fig.  25.    Granular  Casts.     (Greene). 


71 


formations  soluble  in  acetic  acid.  They  are  of  variable  size  and 
generally  difficult  to  detect  on  account  of  their  apparently  struc- 
tureless character.  At  times  a  slight  granulation  may  be  seen 
imbedded  in  or  adhering  to  their  matrix  and  occasionally  acci- 
dental attachments  of  pus  or  fat  globules  in  small  numbers. 

Epithelial  casts  have  a  hyaline  matrix  more  or  less  concealed 
by  epithelial  cells.  The  presence  of  these  casts  is  indicative  of  an 
acute  process. 

Blood  casts  con-  

sist  of  the  hyaline 
matrix  with  blood 
corpuscles  imbed- 
ded in  or  adhering 
to  the  matrix.  Pus 
easts  are  rare, 
but  when  present 
the  pus  corpuscles 
adhere  to  the  mat- 
rix. Blood  casts 
are  indicative  of  a 
hemorrhage  into 
the  tubules  and  of 
an  acute  hemor- 
rhagic process.  Hy- 
aline and  epitheli- 
al casts  are  usual- 
ly associated  with  them. 


Fig.  26.    a.  Fatty  casts,  b.  and  c.  Blood 
casts,    d.  Free  fatty  Molecules.  (Roberts). 


Granular  casts  usually 
have  well  defined  boundaries 
with  granular  matter  im- 
bedded in  or  adhering  to  the 
matrix.  They  may  be  finely 
or  coarsely  granular,  the  lat- 
ter having  a  more  serious 
significance.  Granular  casts 
are  due  to  a  disintegration  of 
FIG.  27.  Fatty  Casts  and  Fat  Droplets,  ^j^^  ^^^^^  epithelium.     Their 

degree  of  refraction  is  changeable;  sometimes  they  appear  yel- 
lowish, at  other  times  colorless. 

Fatty  casts  have  a  hyaline  matrix  containing  a  number   of 


72 

small,  glistening  fat  globules  and  granules.  Some  free  fat  i». 
also  usually  found  in  the  field.  As  fatty  casts  are  secondary 
products  of  epithelial  and  granular  casts,  the  diagnosis  of  a 
chronic  process  is  justifiable.  The  hyaline  matrix  is  character- 
istic of  the  different  varie- 
ties of  casts  that  have- 
been  mentioned  up  to  this 
point. 

Waxy  casts  differ  in 
chemical  composition  from 
those  previously  m  e  n  - 
tioned.  They  are  charac- 
terized by  wavy  contours ; 
a  high  refracting  power;  a 
more  or  less  yellowish 
color  and  quite  a  high  de- 
gree of  brittleness.  They 
FlG.  28.     Waxy  Casts.  ,       ,        -^       -       n       T 

are  slowly,  ir  at  all,  at- 
tacked by  acetic  acid.  Their  presence  signifies  waxy  degenera- 
tion of  the  kidney.  Hyaline  casts  may  sometimes  have  a  super- 
ficial resemblance  to  waxy  casts,  but  they  never  have  the  same 
high  refraction  as  the  latter. 

In  the  urine  of  the  horse,  the  sediment  of  crystals  of  lime  car- 
bonate may  obscure  the  search  for  casts.  In  this  case  add  a  few 
drops  of  acetic  acid  which  will  dissolve  the  crystals  quickly, 
and  the  casts,  if  present,  will  show  more  distinctly  but  undergo 
solution  more  slowly  than  the  crystals. 

Cast-like  formations  are  composed  of  various  elements  having 
a  form  somewhat  similar  to  casts  but  lacking  the  matrix  soluble 
in  acetic  acid.  Amorphous  urates  often  simulate  granular  casts, 
in  form.  Bacteria  are  often  grouped  in  a  manner  similar  to  the 
form  of  a  cast,  but  a  close  inspection  shows  an  irregular  outline, 
and  usually  a  number  of  groupings  not  in  cast  form.  Granular 
detritus  and  hematoidin  may  also  assume  the  form  of  casts ;  like- 
wise epithelial  cells,  blood  corpuscles  and  fibrin  in  renal  hempr- 
rhages  may  also  assume  the  form  of  casts.  Acetic  acid  is  said  to 
be  a  reliable  reagent  for  differentiating  between  true  and  false 
casts.  This  reagent  dissolves  the  matrix  of  the  true  casts  but  does; 
not  act  upon  the  cast-like  formations. 


73 


I 


Cylindroids  appear  like  hyaline  casts,  but  are  large  and 
band-like.  Their  breadth  is  uniform  and  they  often  contain  crys- 
tals, epithelial  cells  and  corpuscles. 
They  are  soluble  in  acetic  acid  and 
are  of  renal  origin.  No  especial  sig- 
nificance is  attached  to  them. 


Mucous  cylinders,  sometimes 
called  cylindroids,  are  usually  not  of 
uniform  breadth  and  seldom  contain 
morphologic  constituents  and  are  in- 
soluble in  acetic  acid.  They  are  usu- 
ally found  in  any  urine  containing 
an  abundance  of  mucus  and  are  of 
no  special  significance. 


Fig.  29.   Cylindroids. 
a.  b,  Cast-like  forms;    c, 
Filamentous    forms.       (Og- 
den). 


Upon  special  blanks,  test  specimens  of  urine  quantitatively, 
the  number  and  amount  of  constituents  being  unknown. 


74 


Form  used  in  examination  of  Horse  Urine. 


No Specie 

es Age 

Sex 

Object 

Date 



Normal  (Horse). 

Sample 

Amount  in  24  hours 

3000-4000  c.  c. 

Specific  Gravity 

1025-1050 

Reaction 

Alkaline 

Color 

Yellowish  brown 

Translucency 

Turbid 

Consistency 

Viscid 

Total  Solids 

50-120  parts  per  1000 

Chlorides 

8-14      '^ 

Sulphates 

2-3 

Phosphates 

.05-.2       " 

Urea 

20-40      '' 

Uric  Acid 

Trace 

Hippuric  Acid 

4-  8  parts  per  1000 

Indican 

.1-.2        '' 

ABNORMAL  CONSTITUENTS. 

Albumin 

Sugar 

Bile 

TTpm  ncl  nViin 

MICROSCOPIC  EXAMINATION. 

Epithelial  Cells 

Leucocytes 

Blood 

Casts 

Spermatozoa 

' 

Micro-organisms 

Crystalline  Deposit. 

Calcium  Carbonate 

Calcium  Oxalate 

Triple  Phosphates 

• 


75 


Form  used  in  Examination  of  Human  Urine. 


• 


iName  

xVClClX  cob 

Date. 

Normal.                          Sample. 

Amount  in  24  hours. 

1000-1500  cc. 

Specific  gravity 

1015-1025 

Reaction 

Acid  (30°-40°) 

Color 

Light  golden 

Translucency 

Clear 

34-50  parts  per  1000 

Solids 

50-75  parts  per  24  hours 

6-9  parts  per  1000  as  NaCl 

Chlorides 

10-15  parts  per  24  hours 

1.5-2.5  parts  per  1000  as  F.O, 

Phosphates 

2.-3.5  parts  per  24  hours 

Sulphates 

1.5-3  parts  per  1000  as  SO^ 

2.5-4  parts  per  24  hours 

Urea 

14-22  parts  per  1000 

22-33  parts  per  24  hours 

Uric  Acid 

0.25-0.40  parts  per  1000 

0.40-0.60  parts  per  24  hours 

Indican 

Normal,  little,  medium,  much 

Albumin 

Sugar 

Bile 

Hemoglobin 

Epithelial  cells 

Leucocytes 

Blood 

Casts 

Crystalline  Deposit. 

Uric  Acid 

Oxalates 

Triple  Phosphates 

Urates 

76  " 

Procedure  in  Examining  a  Sample  of  Urine. 

Experience  has  shown  that  the  following  procedure  ex- 
pedites the  work  of  examination.  Pour  the  urine  into  a  sedi- 
mentation glass,  as  some  little  time  may  be  required  for  the 
sediment  to  settle.  After  noting  the  amount,  color  and  translu- 
cency,  take  the  specific  gravity  with  the  urinometer.  This  may 
be  done  in  the  sedimentation  glass.  Filter  as  much  of  the  urine 
as  may  be  needed  for  the  subsequent  tests.  While  waiting  for 
the  urine  to  filter  make  the  tests  for  urea  and  uric  acid  as  the 
correct  reading  is  not  obtained  until  some  few  minutes  after 
the  tests  have  been  made.  Test  the  reaction  with  litmus  paper 
or  if  the  degree  of  acidity  is  required  (human)  use  the  acidi- 
meter.  Test  for  indican.  Thus  far  the  tests  may  be  made  just  as 
well  with  the  unfiltered  as  the  filtered  urine. 

For  the  albumin  and  sugar  tests  the  filtered  urine  must 
be  used,  using  two  or  three  different  tests  for  each  substance. 
If  albumin  is  present  it  should  be  removed  before  making  the 
sugar  tests.  Also  before  making  the  centrifuge  tests  for  the 
chlorides,  phosphates  and  sulphates.  This  may  be  done  by  add- 
ing a  little  acetic  acid  to  the  urine  and  boiling  it  until  the 
albumin  is  coagulated.  Filter  out  the  albumin  and  use  the 
urine  thus  filtered  for  the  remaining  tests. 

With  a  pipette  remove  some  of  the  sediment  at  the  bottom 
of  the  sedimentation  glass  and  introduce  it  into  a  centrifuge 
tube  filling  the  remainder  of  the  tube  to  the  required  height 
with  the  unfiltered  urine.  Include  this  with  the  other  centri- 
fuge tubes  used  in  making  the  tests  for  the  chlorides,  etc.,  re- 
volving them  in  the  centrifuge  for  three  minutes.  If  albumin  is 
present  and  it  is  desired  to  know  the  amount,  it  may  be  det  r- 
mined  by  the  centrifuge  method  as  described  in  the  text. 

For  the  microscopical  examination,  prepare  a  couple  of 
slides  from  the  sediment  in  the  centrifuge  tube,  after  pouring 
off  the  clear  urine  above.  It  is  well,  also,  to  prepare  a  slide  or 
two  from  the  sediment  in  the  sedimentation  glass.  After  putting 
a  drop  or  two  of  the  sediment  on  the  slide  it  may  be  covered  with 
a  cover  glass  and  examined  clear  or  before  covering  the  sedi- 
ment a  drop  of  dilute  Lugol's  solution,  or  safranin  or  other 
stain  may  be  added.     In  some  cases  the  slight  tint  of  the  dye 


77 

appears  to  make  the  casts  or  other  elements  stand  out  more 
clearly.  To  detect  the  amyloid  bodies  or  glycogen  cells  it  is 
necessary  to  use  the  iodine  solution  and  this  often  times  serves 
to  make  the  casts  and  other  elements  more  distinctly  visible. 


APPENDIX. 


Formulae  for  Reagents. 

Barium  Chloride  Solution  for  Centrifuge  Sulphate  Test. 
Barum  chloride,  4  parts 

Hydrochloric  acid,  1  part 

Distilled  water,  16  parts 

Barium  Chloride  Solution  for  Quantitative  Test  of  Sulphates. 
Pure  crystallized  barium  chloride,  30.5  grams. 

Distilled  water  to  1000.     cc. 

Baryta  Mixture.  This  mixture  is  prepared  by  making  sat- 
urated solutions  in  the  cold,  of  barium  nitrate  and  barium  hy- 
drate, and  adding  two  volumes  of  the  hydrate  to  one  volume  of 
the  nitrate. 

Benedict's  Solution. 


Solution  1.    Cupric  Sulphate 

34.65 

grams 

Distilled  water  up  to 

500. 

cc. 

Solution  2.    Sodium  carbonate   (anhy- 

drous). 

100. 

grams 

Rochelle  salt. 

173. 

grams 

Distilled  water  up  to 

500. 

cc. 

Benedict's  Single  Solution. 

Cupric  Sulphate, 

17.3 

grams 

Sodium  Citrate, 

173. 

grams 

Sodium  Carbonate  (anhydrous). 

100. 

grams 

Distilled  water  up  to 

1000. 

cc. 

Benedict's  Solutions  for  Quantitative  Tests.  For  quantita- 
tive work  three  solutions  are  employed,  each  kept  in  separate  bot- 
tles with  rubber  stoppers  to  prevent  deterioration.    Solution  1  is 


78 

the  cupric  sulphate  solution  as  given  above.  Solution  2  is  the 
alkaline  tartrate  solution  also  given  above.  Solution  3,  the  potas- 
sium ferro-thiocyanate  solution  is  as  follows: 

Potassium  ferrocyanide,  15  grams 

Potassium  thiocyanate,  62.5  grams 

Anhydrous  sodium  carbonate,  50  grams 

Water  up  to  500  cc. 

In  testing  use  equal  volumes  of  the  solutions. 

Cochineal  Solution.  Boil  40  grams  of  cochineal  in  800  cc. 
of  water.    When  cool  add  200  cc.  of  alcohol  and  filter. 

Decinormal  Oxalic  Solution.  Dissolve  6.285  grams  of  pure 
oxalic  acid  in  enough  distilled  water  to  make  at  or  near  15°  C. 
(59°  F.),  exactly  1000  cc. 

Decinormal  Sodium  Hydroxide  Solution.  Dissolve  3.996 
grams  of  pure  sodium  hydroxide  in  enough  distilled  water  to 
make,  at  or  near  15°  C.  (59°  F.),  exactly  1000  cc. 

Ehrlich's  Diazo-reaction. 

Solution  1.    Sulphanilic  acid,  2  grams 

Hydrochloric  acid,  50  cc. 

Distilled  water,  ^      1000  cc. 

Solution  2.  A  0.5%  solution  of  sodium  nitrite.  Use  in  the 
proportion  of  1  part  of  No.  2  to  50  parts  of  No.  1. 

Esbach's  Reagent. 

Picric  acid,  10  grams 

Citric  acid,  20  grams 

Distilled  water  up  to  1000  cc. 

Fehling's  Solution. 

Solution  A.  Pure  Copper  Sulphate  crystalline,  34.64  grams- 
Distilled  water  up  to                       500.  cc. 

Solution  B.  Sodio-potassium  tartrate  (Rochelle 

salt),  173.  grams 

Pure  caustic  potash,  125.  grams 

Distilled  water  up  to  500.  cc. 

Use  equal  parts  of  A  and  B. 


79 

Ferric-Hydrochloric  Acid  Indican  Reagent.  (Obennayer's 
Reagent).  Dissolve  2  grams  of  solid  ferric  chloride  in  500  cc.  of 
concentrated  hydrochloric  acid.    Sp.  gr.  1.19. 

Hypobromite  Solution.  Dissolve  40  grams  of  caustic  soda 
in  100  cc.  of  distilled  water.  To  23  cc.  of  this  solution  add  2  cc. 
of  bromine.  To  this  mixture  add  an  equal  volume  (25  cc.)  of 
water.  The  solution  is  not  very  stable  and  is  best  made  up  •  as 
needed. 

Lugol's  Solution. 

Iodine,  2.5  grams 

Potassium  iodide,  5.     grams 

Distilled  water  to  50.     cc. 

This  may  ordinarily  be  diluted  1  to  5  or  10  for  urine  work. 

Magnesia  Mixture. 

Magnesium  sulphate,  1  part 

Ammonium  chloride,  1  part 

Ammonia  water,  1  part 

Distilled  water,  8  parts 

Millard-Roberts  Reagent. 

Nitric  acid.  1  part 

Sat.  Sol.  Magnesium  Sulphate,         5  parts 

Mohr's  Volumetric  Method  for  Chlorides. 

Solution  1.    Fused  silver  nitrate,  29.075  grams 

Distilled  water  to  make,       1000.       cc. 

Solution  2.    Neutral  potassium  chromate,  10.         grams 
Distilled  water  to  make,         100.         cc. 

Nylander's  Reagent.  Digest  2  grams  of  bismuth  subnitrate 
and  4  grams  of  Rochelle  salt  in  100  cc.  of  a  10%  solution  of 
potassium  hydroxide.  The  reagent  should  then  be  cooled  and  fil- 
tered. 

Phenolphthalein.  Dissolve  1  gram  of  phenolphthalein  in 
100  cc.  of  95%  alcohol. 


30 


Ruhemann's  Uric  Acid  Reagent. 

Iodine,  1.5  grams 

Potassium  iodide,  1.5  grams 

Absolute  alcohol,  15.0  grams 

Distilled  water,  185.0  grams 


Shieb's  Reagent. 

Solution  1.    Ammonium  Sulphate  (purest),    1.2  grams 
Copper  Sulphate  (purest),  2.6  grams 

Distilled  water  up  to  50.     cc. 

Solution  2.    Caustic  potash  C.  P.  20.  grams 

Distilled  water  up  to  50.  cc. 
Dissolve  and  when  cool,  add 

Glycerine,  50.  cc. 

Ammonia  water,  0.960  sp.  gr.  300.  cc. 

Add  No.  1  to  No.  2  and  dilute  the  whole  to  500  cc.  with  dis- 
tilled water.    Stopper  securely  and  shake  until  thoroughly  mixed. 


Noteworthy    Veterinary 
Publications 


The  Pathology  and  Differential  Diagnosis  of  Infectious  Diseases  of 

Animals.     Moore. $4.00 

General  Surgery — Frohner.     Translated  by  Udall. 3.00 

Veterinary  Doses  and  Prescription  Writing.     Fish 1.50 

Examination  of  the  Urine  of  the  Horse  and  Man.     Fish. 1.50 

The  Clinical  Pathology    of    the   Blood    of    Domesticated   Animals 

Burnett. 2.50 

Surgical  and  Obstetrical  Operations.     Williams. 2.50 

Pure  Milk  and  the    Public    Health.     Ward. 2.00 

Veterinary  Medicine.     Law.     Vol.  I,  II,  III.  IV,  V.  Per  vol. 4.00 

Veterinary  Obstetrics.     Williams. 8.00 

Exercises  in  Materia    Medica  and  Pharmacy.     Fish. 1.50 

Exercises  in  Physiology.     Fish 1.50 


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CARPENTER  <&,  CO. 

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JAN  1 1  2000 

^^B  2  3  2001 
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